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Rosa

Rosa: A versatile genus of flowering plants in the family Rosaceae, known for their beautiful and fragrant blooms.
These shrubs are widely cultivated for ornamental purposes, with a diverse array of species and cultivars offering a range of colors, forms, and growth habits.
Roses are often used in landscaping, cut flower arrangements, and perfume production.
They can be found growing in temperate regions around the world and have a long history of cultural significance.
Researchers may leverage Rosa species and cultivars in a variety of applications, such as the study of plant biology, the development of novel horticultural traits, or the extraction of valuable compounds.
This MeSH term provides a concise overview of the Rosa genus and its potential research applications.

Most cited protocols related to «Rosa»

1
Simulating 16S Sequence Alignments
The simulated protein alignments and the genuine COG alignments were described previously [2] (link). The 16S alignment with 237,882 distinct sequences was taken from GreenGenes [33] (link) (http://greengenes.lbl.gov). The 16S alignment with 15,011 distinct “families” is a non-redundant subset of these sequences ( identical). 16S alignments with 500 sequences are also non-redundant random subsets ( identical). Other large 16S alignments are from [11] (link).
For the 16S-like simulations with 78,132 distinct sequences, we used a maximum-likelihood tree inferred from a non-redundant aligned subset of the full set of 16S sequences ( % identity) by an earlier version of FastTree (1.9) with the Jukes-Cantor model (no CAT). To ensure that the simulated trees were resolvable, which facilitates comparison of methods (but inflates the accuracy of all methods), branch lengths of less than 0.001 were replaced with values of 0.001, which corresponds to roughly one substitution across the internal branch, as the 16S alignment has 1,287 positions. Evolutionary rates for each site were randomly selected from 16 rate categories according to a gamma distribution with a coefficient of variation of 0.7. Given the tree and the rates, sequences were simulated with Rose [34] (link) under the HKY model and no transition bias. To allow Rose to handle branch lengths of less than 1%, we set “MeanSubstitution = 0.00134” and multiplied the branch lengths by 1,000.
Price M.N., Dehal P.S, & Arkin A.P. (2010). FastTree 2 – Approximately Maximum-Likelihood Trees for Large Alignments. PLoS ONE, 5(3), e9490.
Full text: Click here
Publication 2010
Biological Evolution Cantor Gamma Rays Proteins Sequence Alignment Trees
2
Simulated Evolutionary Alignments Generator
We obtained sequences of members of Clusters of Orthologous Groups (COG) gene families (Tatusov et al. 2001) and members of Pfam PF00005 (Finn et al. 2006 ) from the fall 2007 release of the MicrobesOnline database (http://www.microbesonline.org/). We aligned the sequences to the family's profile, using reverse position-specific Blast for the COG alignment (Schaffer et al. 2001 (link)) and hmmalign for the PF00005 alignment (http://hmmer.janelia.org/). As the profiles only include positions that are present in many members of the family, these alignments do not contain all positions from the original sequences. The 16S rRNA alignment is from greengenes and is trimmed with the greengenes mask (DeSantis et al. 2006 (link); http://greengenes.lbl.gov).
To simulate alignments with realistic phylogenies and realistic gaps, we used the COG alignments. In each simulation, we selected the desired number of sequences from a COG alignment, we removed positions that were over 25% gaps, we estimated a topology and branch lengths with PhyML (Guindon and Gascuel 2003 (link)), we estimated evolutionary rates across sites with PHYLIP's proml (http://evolution.genetics.washington.edu/phylip.htm), we simulated sequences with Rose (Stoye et al. 1998 (link)), and we reintroduced the gaps from the original alignment. For simulations of 5,000 sequences, we used FastTree instead of PhyML and we assigned evolutionary rates at random. For N = 10, we simulated 3,100 alignments (10 independent runs per family); for N = 50, we simulated 3,099 alignments; for N = 250, we simulated 308 alignments; for N = 1,250, we simulated only 92 alignments because some PhyML jobs did not complete, and for N = 5,000, we simulated 7 alignments, as only seven families contained enough nonredundant sequences. See supplementary note 2 (Supplementary Material online) for technical details.
Price M.N., Dehal P.S, & Arkin A.P. (2009). FastTree: Computing Large Minimum Evolution Trees with Profiles instead of a Distance Matrix. Molecular Biology and Evolution, 26(7), 1641-1650.
Publication 2009
Biological Evolution Family Member Genes RNA, Ribosomal, 16S
3
Robust Knock-in Mouse Line Generation
Targeting constructs were generated using a combined gene synthesis (GenScript Corp.) and molecular cloning approach. Briefly, to target the Rosa26 locus, a cassette containing the following components was constructed: FRT – LoxP – Stop codons – 3x SV40 polyA – LoxP – EYFP – WPRE – bGH polyA – AttB – PGK promoter – FRT – Neo – PGK polyA – AttP. For most targeting vectors, this cassette was cloned into a Rosa-CAG targeting vector3 (link), downstream of the CAG promoter and upstream of the 3′ arm, to generate the final EYFP targeting vector. Unique restriction sites flanking the EYFP gene were used to replace EYFP with alternative reporter genes. For the Ai2 vector, which lacks the WPRE, the CAG promoter was inserted between the first FRT and LoxP sites, and the cassette was cloned immediately downstream of the 5′ homology arm. The final targeting vectors contained 5′ and 3′ homology arms of 1.1 kb and 4.3 kb, as well as a PGK-DTA cassette for negative selection. Targeting constructs for knock-in Cre lines inserted into other gene loci were constructed in similar ways.
The targeting vectors were linearized and transfected into the 129/B6 F1 hybrid ES cell line G442 (link) using an Amaxa electroporator. G418-resistant ES clones were screened by Southern blot analysis of HindIII digested DNA, which was probed with a 1.1 kb genomic fragment from immediately upstream of the 5′ arm. We observed a recombination rate of about 25% for the four constructs. Positive ES clones were injected into C57BL/6J blastocysts to obtain chimeric mice following standard procedures. Both ES cell transfections and blastocyst injections were performed by the University of Washington Transgenic Resources Program. Due to the robustness of the G4 cells, high-percentage chimeras and high rates of germline transmission were routinely obtained. Chimeric mice were bred with either C57BL/6J mice to obtain germline transmission or various Cre-driver lines for direct characterization.
An Ai9 ES cell clone with strong germline transmission potency was used in subsequent transfections for the Flp-mediated exchange strategy outlined in Supplementary Figure 4 online. Ai9 ES cells were co-transfected using a Bio-Rad electroporator with 100 μg of pCAGGS-FLPe (Open Biosystems) and 40 μg of an incoming replacement vector. After 8 to 10 days of Hygromycin B selection, surviving colonies that also appeared green by fluorescence microscopy were picked and screened by PCR using primer sets designed to confirm a correct insertion of the incoming vector at the 5′ and 3′ FRT recombinase sites.
Madisen L., Zwingman T.A., Sunkin S.M., Oh S.W., Zariwala H.A., Gu H., Ng L.L., Palmiter R.D., Hawrylycz M.J., Jones A.R., Lein E.S, & Zeng H. (2009). A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nature neuroscience, 13(1), 133-140.
Publication 2009
Anabolism Animals, Transgenic antibiotic G 418 Blastocyst Cells Chimera Clone Cells Cloning Vectors Codon, Terminator Embryonic Stem Cells Genes Genes, Reporter Genetic Loci Genome Germ Cells Germ Line Hybrid Cells Hygromycin B Mice, Inbred C57BL Microscopy, Fluorescence Mus N-fluoresceinylphosphatidylethanolamine Oligonucleotide Primers Poly A Recombinase Recombination, Genetic Rosa Simian virus 40 Southern Blotting Transfection Transmission, Communicable Disease
4
Robust Knock-in Mouse Line Generation
Targeting constructs were generated using a combined gene synthesis (GenScript Corp.) and molecular cloning approach. Briefly, to target the Rosa26 locus, a cassette containing the following components was constructed: FRT – LoxP – Stop codons – 3x SV40 polyA – LoxP – EYFP – WPRE – bGH polyA – AttB – PGK promoter – FRT – Neo – PGK polyA – AttP. For most targeting vectors, this cassette was cloned into a Rosa-CAG targeting vector3 (link), downstream of the CAG promoter and upstream of the 3′ arm, to generate the final EYFP targeting vector. Unique restriction sites flanking the EYFP gene were used to replace EYFP with alternative reporter genes. For the Ai2 vector, which lacks the WPRE, the CAG promoter was inserted between the first FRT and LoxP sites, and the cassette was cloned immediately downstream of the 5′ homology arm. The final targeting vectors contained 5′ and 3′ homology arms of 1.1 kb and 4.3 kb, as well as a PGK-DTA cassette for negative selection. Targeting constructs for knock-in Cre lines inserted into other gene loci were constructed in similar ways.
The targeting vectors were linearized and transfected into the 129/B6 F1 hybrid ES cell line G442 (link) using an Amaxa electroporator. G418-resistant ES clones were screened by Southern blot analysis of HindIII digested DNA, which was probed with a 1.1 kb genomic fragment from immediately upstream of the 5′ arm. We observed a recombination rate of about 25% for the four constructs. Positive ES clones were injected into C57BL/6J blastocysts to obtain chimeric mice following standard procedures. Both ES cell transfections and blastocyst injections were performed by the University of Washington Transgenic Resources Program. Due to the robustness of the G4 cells, high-percentage chimeras and high rates of germline transmission were routinely obtained. Chimeric mice were bred with either C57BL/6J mice to obtain germline transmission or various Cre-driver lines for direct characterization.
An Ai9 ES cell clone with strong germline transmission potency was used in subsequent transfections for the Flp-mediated exchange strategy outlined in Supplementary Figure 4 online. Ai9 ES cells were co-transfected using a Bio-Rad electroporator with 100 μg of pCAGGS-FLPe (Open Biosystems) and 40 μg of an incoming replacement vector. After 8 to 10 days of Hygromycin B selection, surviving colonies that also appeared green by fluorescence microscopy were picked and screened by PCR using primer sets designed to confirm a correct insertion of the incoming vector at the 5′ and 3′ FRT recombinase sites.
Madisen L., Zwingman T.A., Sunkin S.M., Oh S.W., Zariwala H.A., Gu H., Ng L.L., Palmiter R.D., Hawrylycz M.J., Jones A.R., Lein E.S, & Zeng H. (2009). A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nature neuroscience, 13(1), 133-140.
Publication 2009
Anabolism Animals, Transgenic antibiotic G 418 Blastocyst Cells Chimera Clone Cells Cloning Vectors Codon, Terminator Embryonic Stem Cells Genes Genes, Reporter Genetic Loci Genome Germ Cells Germ Line Hybrid Cells Hygromycin B Mice, Inbred C57BL Microscopy, Fluorescence Mus N-fluoresceinylphosphatidylethanolamine Oligonucleotide Primers Poly A Recombinase Recombination, Genetic Rosa Simian virus 40 Southern Blotting Transfection Transmission, Communicable Disease
5
Improved Protein Secondary Structure Prediction
As with JPred3, JPred4 makes secondary structure and residue solvent accessibility predictions by the JNet algorithm (11 (link),31 (link)). However, in JPred4, the JNet 2.0 neural network-based predictor has been retrained to make JNet 2.3.1 by 7-fold cross-validation using one representative for each of the 1358 SCOPe/ASTRAL v.2.04 superfamily domain sequences (32 (link)). Multiple alignments for each sequence were built by PSI-BLAST (33 (link)) through searching UniRef90 v.2014_07 (34 (link)). In addition to retraining, the HMM building step in JNet was updated to HMMer 3 (35 (link)) and some improvements were made to the code to simplify management and future algorithmic developments. The final accuracy of JNet 2.3.1 was assessed in a blind test on 150 sequences from 150 superfamilies not used in training. The 150 superfamily sequences were selected to reproduce a similar distribution of secondary structure compositions as the training structures in order to avoid biasing the reported accuracy of the blind test results. On the blind test, the average secondary structure prediction Q3 score increased to 82.0% from 81.5% for JNet v.2.0, and solvent accessibility prediction accuracy rose to 90.0, 83.6 and 78.1% from 88.9, 82.4 and 77.8% for JNet v.2.0 for each of >0, >5 and >25% relative solvent accessibility thresholds.
Drozdetskiy A., Cole C., Procter J, & Barton G.J. (2015). JPred4: a protein secondary structure prediction server. Nucleic Acids Research, 43(Web Server issue), W389-W394.
Full text: Click here
Publication 2015
Aster Plant Sequence Alignment Solvents Visually Impaired Persons

Most recents protocols related to «Rosa»

1
Microbial Seed Treatments for Enhanced Crop Performance
Not available on PMC !

Example 2

A. Seed Treatment with Isolated Microbe

In this example, an isolated microbe from Tables 1-3 will be applied as a seed coating to seeds of corn (Zea mays). Upon applying the isolated microbe as a seed coating, the corn will be planted and cultivated in the standard manner.

A control plot of corn seeds, which did not have the isolated microbe applied as a seed coating, will also be planted.

It is expected that the corn plants grown from the seeds treated with the seed coating will exhibit a quantifiably higher biomass than the control corn plants.

The biomass from the treated plants may be about 1-10% higher, 10-20% higher, 20-30% higher, 30-40% higher, 40-50% higher, 50-60% higher, 60-70% higher, 70-80% higher, 80-90% higher, or more.

The biomass from the treated plants may equate to about a 1 bushel per acre increase over the controls, or a 2 bushel per acre increase, or a 3 bushel per acre increase, or a 4 bushel per acre increase, or a 5 bushel per acre increase, or more.

In some aspects, the biomass increase is statistically significant. In other aspects, the biomass increase is not statistically significant, but is still quantifiable.

B. Seed Treatment with Microbial Consortia

In this example, a microbial consortium, comprising at least two microbes from Tables 1-3 will be applied as a seed coating to seeds of corn (Zea mays). Upon applying the microbial consortium as a seed coating, the corn will be planted and cultivated in the standard manner.

A control plot of corn seeds, which did not have the microbial consortium applied as a seed coating, will also be planted.

It is expected that the corn plants grown from the seeds treated with the seed coating will exhibit a quantifiably higher biomass than the control corn plants.

The biomass from the treated plants may be about 1-10% higher, 10-20% higher, 20-30% higher, 30-40% higher, 40-50% higher, 50-60% higher, 60-70% higher, 70-80% higher, 80-90% higher, or more.

The biomass from the treated plants may equate to about a 1 bushel per acre increase over the controls, or a 2 bushel per acre increase, or a 3 bushel per acre increase, or a 4 bushel per acre increase, or a 5 bushel per acre increase, or more.

In some aspects, the biomass increase is statistically significant. In other aspects, the biomass increase is not statistically significant, but is still quantifiable.

C. Treatment with Agricultural Composition Comprising Isolated Microbe

In this example, an isolated microbe from Tables 1-3 will be applied as an agricultural composition, administered to the corn seed at the time of sowing.

For example, it is anticipated that a farmer will apply the agricultural composition to the corn seeds simultaneously upon planting the seeds into the field. This can be accomplished, for example, by applying the agricultural composition to a hopper/bulk tank on a standard 16 row planter, which contains the corn seeds and which is configured to plant the same into rows. Alternatively, the agricultural composition can be contained in a separate bulk tank on the planter and sprayed into the rows upon planting the corn seed.

A control plot of corn seeds, which are not administered the agricultural composition, will also be planted.

It is expected that the corn plants grown from the seeds treated with the agricultural composition will exhibit a quantifiably higher biomass than the control corn plants.

The biomass from the treated plants may be about 1-10% higher, 10-20% higher, 20-30% higher, 30-40% higher, 40-50% higher, 50-60% higher, 60-70% higher, 70-80% higher, 80-90% higher, or more.

The biomass from the treated plants may equate to about a 1 bushel per acre increase over the controls, or a 2 bushel per acre increase, or a 3 bushel per acre increase, or a 4 bushel per acre increase, or a 5 bushel per acre increase, or more.

In some aspects, the biomass increase is statistically significant. In other aspects, the biomass increase is not statistically significant, but is still quantifiable.

D. Treatment with Agricultural Composition Comprising Microbial Consortia

In this example, a microbial consortium, comprising at least two microbes from Tables 1-3 will be applied as an agricultural composition, administered to the corn seed at the time of sowing.

For example, it is anticipated that a farmer will apply the agricultural composition to the corn seeds simultaneously upon planting the seeds into the field. This can be accomplished, for example, by applying the agricultural composition to a hopper/bulk tank on a standard 16 row planter, which contains the corn seeds and which is configured to plant the same into rows. Alternatively, the agricultural composition can be contained in a separate bulk tank on the planter and sprayed into the rows upon planting the corn seed.

A control plot of corn seeds, which are not administered the agricultural composition, will also be planted.

It is expected that the corn plants grown from the seeds treated with the agricultural composition will exhibit a quantifiably higher biomass than the control corn plants.

The biomass from the treated plants may be about 1-10% higher, 10-20% higher, 20-30% higher, 30-40% higher, 40-50% higher, 50-60% higher, 60-70% higher, 70-80% higher, 80-90% higher, or more.

The biomass from the treated plants may equate to about a 1 bushel per acre increase over the controls, or a 2 bushel per acre increase, or a 3 bushel per acre increase, or a 4 bushel per acre increase, or a 5 bushel per acre increase, or more.

In some aspects, the biomass increase is statistically significant. In other aspects, the biomass increase is not statistically significant, but is still quantifiable.

A. Seed Treatment with Isolated Microbe

In this example, an isolated microbe from Tables 1-3 will be applied as a seed coating to seeds of corn (Zea mays). Upon applying the isolated microbe as a seed coating, the corn will be planted and cultivated in the standard manner.

A control plot of corn seeds, which did not have the isolated microbe applied as a seed coating, will also be planted.

It is expected that the corn plants grown from the seeds treated with the seed coating will exhibit a quantifiable and superior ability to tolerate drought conditions and/or exhibit superior water use efficiency, as compared to the control corn plants.

The drought tolerance and/or water use efficiency can be based on any number of standard tests from the art, e.g leaf water retention, turgor loss point, rate of photosynthesis, leaf color and other phenotypic indications of drought stress, yield performance, and various root morphological and growth patterns.

B. Seed Treatment with Microbial Consortia

In this example, a microbial consortium, comprising at least two microbes from Tables 1-3 will be applied as a seed coating to seeds of corn (Zea mays). Upon applying the microbial consortium as a seed coating, the corn will be planted and cultivated in the standard manner.

A control plot of corn seeds, which did not have the microbial consortium applied as a seed coating, will also be planted.

It is expected that the corn plants grown from the seeds treated with the seed coating will exhibit a quantifiable and superior ability to tolerate drought conditions and/or exhibit superior water use efficiency, as compared to the control corn plants.

The drought tolerance and/or water use efficiency can be based on any number of standard tests from the art, e.g leaf water retention, turgor loss point, rate of photosynthesis, leaf color and other phenotypic indications of drought stress, yield performance, and various root morphological and growth patterns.

C. Treatment with Agricultural Composition Comprising Isolated Microbe

In this example, an isolated microbe from Tables 1-3 will be applied as an agricultural composition, administered to the corn seed at the time of sowing.

For example, it is anticipated that a farmer will apply the agricultural composition to the corn seeds simultaneously upon planting the seeds into the field. This can be accomplished, for example, by applying the agricultural composition to a hopper/bulk tank on a standard 16 row planter, which contains the corn seeds and which is configured to plant the same into rows. Alternatively, the agricultural composition can be contained in a separate bulk tank on the planter and sprayed into the rows upon planting the corn seed.

A control plot of corn seeds, which are not administered the agricultural composition, will also be planted.

It is expected that the corn plants grown from the seeds treated with the with the agricultural composition will exhibit a quantifiable and superior ability to tolerate drought conditions and/or exhibit superior water use efficiency, as compared to the control corn plants.

The drought tolerance and/or water use efficiency can be based on any number of standard tests from the art, e.g leaf water retention, turgor loss point, rate of photosynthesis, leaf color and other phenotypic indications of drought stress, yield performance, and various root morphological and growth patterns.

D. Treatment with Agricultural Composition Comprising Microbial Consortia

In this example, a microbial consortium, comprising at least two microbes from Tables 1-3 will be applied as an agricultural composition, administered to the corn seed at the time of sowing.

For example, it is anticipated that a farmer will apply the agricultural composition to the corn seeds simultaneously upon planting the seeds into the field. This can be accomplished, for example, by applying the agricultural composition to a hopper/bulk tank on a standard 16 row planter, which contains the corn seeds and which is configured to plant the same into rows. Alternatively, the agricultural composition can be contained in a separate bulk tank on the planter and sprayed into the rows upon planting the corn seed.

A control plot of corn seeds, which are not administered the agricultural composition, will also be planted.

It is expected that the corn plants grown from the seeds treated with the with the agricultural composition will exhibit a quantifiable and superior ability to tolerate drought conditions and/or exhibit superior water use efficiency, as compared to the control corn plants.

The drought tolerance and/or water use efficiency can be based on any number of standard tests from the art, e.g leaf water retention, turgor loss point, rate of photosynthesis, leaf color and other phenotypic indications of drought stress, yield performance, and various root morphological and growth patterns.

A. Seed Treatment with Isolated Microbe

In this example, an isolated microbe from Tables 1-3 will be applied as a seed coating to seeds of corn (Zea mays). Upon applying the isolated microbe as a seed coating, the corn will be planted and cultivated in the standard manner.

A control plot of corn seeds, which did not have the isolated microbe applied as a seed coating, will also be planted.

It is expected that the corn plants grown from the seeds treated with the seed coating will exhibit a quantifiable and superior ability to utilize nitrogen, as compared to the control corn plants.

The nitrogen use efficiency can be quantified by recording a measurable change in any of the main nitrogen metabolic pool sizes in the assimilation pathways (e.g., a measurable change in one or more of the following: nitrate, nitrite, ammonia, glutamic acid, aspartic acid, glutamine, asparagine, lysine, leucine, threonine, methionine, glycine, tryptophan, tyrosine, total protein content of a plant part, total nitrogen content of a plant part, and/or chlorophyll content), or where the treated plant is shown to provide the same or elevated biomass or harvestable yield at lower nitrogen fertilization levels compared to the control plant, or where the treated plant is shown to provide elevated biomass or harvestable yields at the same nitrogen fertilization levels compared to a control plant.

B. Seed Treatment with Microbial Consortia

In this example, a microbial consortium, comprising at least two microbes from Tables 1-3 will be applied as a seed coating to seeds of corn (Zea mays). Upon applying the microbial consortium as a seed coating, the corn will be planted and cultivated in the standard manner.

A control plot of corn seeds, which did not have the microbial consortium applied as a seed coating, will also be planted.

It is expected that the corn plants grown from the seeds treated with the seed coating will exhibit a quantifiable and superior ability to utilize nitrogen, as compared to the control corn plants.

The nitrogen use efficiency can be quantified by recording a measurable change in any of the main nitrogen metabolic pool sizes in the assimilation pathways (e.g., a measurable change in one or more of the following: nitrate, nitrite, ammonia, glutamic acid, aspartic acid, glutamine, asparagine, lysine, leucine, threonine, methionine, glycine, tryptophan, tyrosine, total protein content of a plant part, total nitrogen content of a plant part, and/or chlorophyll content), or where the treated plant is shown to provide the same or elevated biomass or harvestable yield at lower nitrogen fertilization levels compared to the control plant, or where the treated plant is shown to provide elevated biomass or harvestable yields at the same nitrogen fertilization levels compared to a control plant.

C. Treatment with Agricultural Composition Comprising Isolated Microbe

In this example, an isolated microbe from Tables 1-3 will be applied as an agricultural composition, administered to the corn seed at the time of sowing.

For example, it is anticipated that a farmer will apply the agricultural composition to the corn seeds simultaneously upon planting the seeds into the field. This can be accomplished, for example, by applying the agricultural composition to a hopper/bulk tank on a standard 16 row planter, which contains the corn seeds and which is configured to plant the same into rows. Alternatively, the agricultural composition can be contained in a separate bulk tank on the planter and sprayed into the rows upon planting the corn seed.

A control plot of corn seeds, which are not administered the agricultural composition, will also be planted.

It is expected that the corn plants grown from the seeds treated with the agricultural composition will exhibit a quantifiable and superior ability to utilize nitrogen, as compared to the control corn plants.

The nitrogen use efficiency can be quantified by recording a measurable change in any of the main nitrogen metabolic pool sizes in the assimilation pathways (e.g., a measurable change in one or more of the following: nitrate, nitrite, ammonia, glutamic acid, aspartic acid, glutamine, asparagine, lysine, leucine, threonine, methionine, glycine, tryptophan, tyrosine, total protein content of a plant part, total nitrogen content of a plant part, and/or chlorophyll content), or where the treated plant is shown to provide the same or elevated biomass or harvestable yield at lower nitrogen fertilization levels compared to the control plant, or where the treated plant is shown to provide elevated biomass or harvestable yields at the same nitrogen fertilization levels compared to a control plant.

D. Treatment with Agricultural Composition Comprising Microbial Consortia

In this example, a microbial consortium, comprising at least two microbes from Tables 1-3 will be applied as an agricultural composition, administered to the corn seed at the time of sowing.

For example, it is anticipated that a farmer will apply the agricultural composition to the corn seeds simultaneously upon planting the seeds into the field. This can be accomplished, for example, by applying the agricultural composition to a hopper/bulk tank on a standard 16 row planter, which contains the corn seeds and which is configured to plant the same into rows. Alternatively, the agricultural composition can be contained in a separate bulk tank on the planter and sprayed into the rows upon planting the corn seed.

A control plot of corn seeds, which are not administered the agricultural composition, will also be planted.

It is expected that the corn plants grown from the seeds treated with the agricultural composition will exhibit a quantifiable and superior ability to utilize nitrogen, as compared to the control corn plants.

The nitrogen use efficiency can be quantified by recording a measurable change in any of the main nitrogen metabolic pool sizes in the assimilation pathways (e.g., a measurable change in one or more of the following: nitrate, nitrite, ammonia, glutamic acid, aspartic acid, glutamine, asparagine, lysine, leucine, threonine, methionine, glycine, tryptophan, tyrosine, total protein content of a plant part, total nitrogen content of a plant part, and/or chlorophyll content), or where the treated plant is shown to provide the same or elevated biomass or harvestable yield at lower nitrogen fertilization levels compared to the control plant, or where the treated plant is shown to provide elevated biomass or harvestable yields at the same nitrogen fertilization levels compared to a control plant.

The inoculants were prepared from isolates grown as spread plates on R2A incubated at 25° C. for 48 to 72 hours. Colonies were harvested by blending with sterile distilled water (SDW) which was then transferred into sterile containers. Serial dilutions of the harvested cells were plated and incubated at 25° C. for 24 hours to estimate the number of colony forming units (CFU) in each suspension. Dilutions were prepared using individual isolates or blends of isolates (consortia) to deliver 1×105 cfu/microbe/seed and seeds inoculated by either imbibition in the liquid suspension or by overtreatment with 5% vegetable gum and oil.

Seeds corresponding to the plants of table 15 were planted within 24 to 48 hours of treatment in agricultural soil, potting media or inert growing media. Plants were grown in small pots (28 mL to 200 mL) in either a controlled environment or in a greenhouse. Chamber photoperiod was set to 16 hours for all experiments on all species. Air temperature was typically maintained between 22-24° C.

Unless otherwise stated, all plants were watered with tap water 2 to 3 times weekly. Growth conditions were varied according to the trait of interest and included manipulation of applied fertilizer, watering regime and salt stress as follows:

    • Low N—seeds planted in soil potting media or inert growing media with no applied N fertilizer
    • Moderate N—seeds planted in soil or growing media supplemented with commercial N fertilizer to equivalent of 135 kg/ha applied N
    • Insol P—seeds planted in potting media or inert growth substrate and watered with quarter strength Pikovskaya's liquid medium containing tri-calcium phosphate as the only form phosphate fertilizer.
    • Cold Stress—seeds planted in soil, potting media or inert growing media and incubated at 10° C. for one week before being transferred to the plant growth room.
    • Salt stress—seeds planted in soil, potting media or inert growing media and watered with a solution containing between 100 to 200 mg/L NaCl.

Untreated (no applied microbe) controls were prepared for each experiment. Plants were randomized on trays throughout the growth environment. Between 10 and 30 replicate plants were prepared for each treatment in each experiment. Phenotypes were measured during early vegetative growth, typically before the V3 developmental stage and between 3 and 6 weeks after sowing. Foliage was cut and weighed. Roots were washed, blotted dry and weighed. Results indicate performance of treatments against the untreated control.

TABLE 15
StrainShootRoot
Microbe sp.IDCropAssayIOC (%)IOC (%)
Bosea thiooxidans123EfficacyEfficacy
overall100%100%
Bosea thiooxidans54522WheatEarly vigor - insol P30-40 —
Bosea thiooxidans54522RyegrassEarly vigor50-60 50-60 
Bosea thiooxidans54522RyegrassEarly vigor - moderate P0-100-10
Duganella violaceinigra111EfficacyEfficacy
overall100%100%
Duganella violaceinigra66361TomatoEarly vigor0-100-10
Duganella violaceinigra66361TomatoEarly vigor30-40 40-50 
Duganella violaceinigra66361TomatoEarly vigor20-30 20-30 
Herbaspirillum huttiense222Efficacy—
overall100%
Herbaspirillum huttiense54487WheatEarly vigor - insol P30-40 —
Herbaspirillum huttiense60507MaizeEarly vigor - salt stress0-100-10
Janthinobacterium sp.222Efficacy—
Overall100%
Janthinobacterium sp.54456WheatEarly vigor - insol P30-40 —
Janthinobacterium sp.54456WheatEarly vigor - insol P0-10—
Janthinobacterium sp.63491RyegrassEarly vigor - drought0-100-10
stress
Massilia niastensis112EfficacyEfficacy
overall80%80%
Massilia niastensis55184WheatEarly vigor - salt stress0-1020-30 
Massilia niastensis55184WinterEarly vigor - cold stress0-1010-20 
wheat
Massilia niastensis55184WinterEarly vigor - cold stress20-30 20-30 
wheat
Massilia niastensis55184WinterEarly vigor - cold stress10-20 10-20 
wheat
Massilia niastensis55184WinterEarly vigor - cold stress<0<0
wheat
Novosphingobium rosa211EfficacyEfficacy
overall100%100%
Novosphingobium rosa65589MaizeEarly vigor - cold stress0-100-10
Novosphingobium rosa65619MaizeEarly vigor - cold stress0-100-10
Paenibacillus amylolyticus111EfficacyEfficacy
overall100%100%
Paenibacillus amylolyticus66316TomatoEarly vigor0-100-10
Paenibacillus amylolyticus66316TomatoEarly vigor10-20 10-20 
Paenibacillus amylolyticus66316TomatoEarly vigor0-100-10
Pantoea agglomerans323EfficacyEfficacy
33%50%
Pantoea agglomerans54499WheatEarly vigor - insol P40-50 —
Pantoea agglomerans57547MaizeEarly vigor - low N<00-10
Pantoea vagans55529MaizeEarly vigor<0<0
(formerly P. agglomerans)
Polaromonas ginsengisoli111EfficacyEfficacy
66%100%
Polaromonas ginsengisoli66373TomatoEarly vigor0-100-10
Polaromonas ginsengisoli66373TomatoEarly vigor20-30 30-40 
Polaromonas ginsengisoli66373TomatoEarly vigor<010-20 
Pseudomonas fluorescens122Efficacy—
100%
Pseudomonas fluorescens54480WheatEarly vigor - insol P>100 —
Pseudomonas fluorescens56530MaizeEarly vigor - moderate N0-10—
Rahnella aquatilis334EfficacyEfficacy
80%63%
Rahnella aquatilis56532MaizeEarly vigor - moderate N10-20 —
Rahnella aquatilis56532MaizeEarly vigor - moderate N0-100-10
Rahnella aquatilis56532WheatEarly vigor - cold stress0-1010-20 
Rahnella aquatilis56532WheatEarly vigor - cold stress<00-10
Rahnella aquatilis56532WheatEarly vigor - cold stress10-20 <0
Rahnella aquatilis57157RyegrassEarly vigor<0—
Rahnella aquatilis57157MaizeEarly vigor - low N0-100-10
Rahnella aquatilis57157MaizeEarly vigor - low N0-10<0
Rahnella aquatilis58013MaizeEarly vigor0-1010-20 
Rahnella aquatilis58013MaizeEarly vigor - low N0-10<0
Rhodococcus erythropolis313Efficacy—
66%
Rhodococcus erythropolis54093MaizeEarly vigor - low N40-50 —
Rhodococcus erythropolis54299MaizeEarly vigor - insol P>100 —
Rhodococcus erythropolis54299MaizeEarly vigor<0<0
Stenotrophomonas chelatiphaga611EfficacyEfficacy
60%60%
Stenotrophomonas chelatiphaga54952MaizeEarly vigor0-100-10
Stenotrophomonas chelatiphaga47207MaizeEarly vigor<0 0
Stenotrophomonas chelatiphaga64212MaizeEarly vigor0-1010-20 
Stenotrophomonas chelatiphaga64208MaizeEarly vigor0-100-10
Stenotrophomonas chelatiphaga58264MaizeEarly vigor<0<0
Stenotrophomonas maltophilia612EfficacyEfficacy
43%66%
Stenotrophomonas maltophilia54073MaizeEarly vigor - low N50-60 —
Stenotrophomonas maltophilia54073MaizeEarly vigor<00-10
Stenotrophomonas maltophilia56181MaizeEarly vigor0-10<0
Stenotrophomonas maltophilia54999MaizeEarly vigor0-100-10
Stenotrophomonas maltophilia54850MaizeEarly vigor 00-10
Stenotrophomonas maltophilia54841MaizeEarly vigor<00-10
Stenotrophomonas maltophilia46856MaizeEarly vigor<0<0
Stenotrophomonas rhizophila811EfficacyEfficacy
12.5%37.5%
Stenotrophomonas rhizophila50839MaizeEarly vigor<0<0
Stenotrophomonas rhizophila48183MaizeEarly vigor<0<0
Stenotrophomonas rhizophila45125MaizeEarly vigor<0<0
Stenotrophomonas rhizophila46120MaizeEarly vigor<00-10
Stenotrophomonas rhizophila46012MaizeEarly vigor<0<0
Stenotrophomonas rhizophila51718MaizeEarly vigor0-100-10
Stenotrophomonas rhizophila66478MaizeEarly vigor<0<0
Stenotrophomonas rhizophila65303MaizeEarly vigor<00-10
Stenotrophomonas terrae221EfficacyEfficacy
50%50%
Stenotrophomonas terrae68741MaizeEarly vigor<0<0
Stenotrophomonas terrae68599MaizeEarly vigor<00-10
Stenotrophomonas terrae68599Capsicum *Early vigor20-30 20-30 
Stenotrophomonas terrae68741Capsicum *Early vigor10-20 20-30 

The data presented in table 15 describes the efficacy with which a microbial species or strain can change a phenotype of interest relative to a control run in the same experiment. Phenotypes measured were shoot fresh weight and root fresh weight for plants growing either in the absence of presence of a stress (assay). For each microbe species, an overall efficacy score indicates the percentage of times a strain of that species increased a both shoot and root fresh weight in independent evaluations. For each species, the specifics of each independent assay is given, providing a strain ID (strain) and the crop species the assay was performed on (crop). For each independent assay the percentage increase in shoot and root fresh weight over the controls is given.

US11871752B2. Agriculturally beneficial microbes, microbial compositions, and consortia (2024-01-16). BIOCONSORTIA, INC. [US]. Inventors: Peter Wigley [NZ], Susan Turner [US], Caroline George [NZ], Thomas Williams [US], Kelly Roberts [US], Graham Hymus [US], Kelvin Lau [NZ].
Full text: Click here
Patent 2024
Ammonia Asparagine Aspartic Acid Biological Assay Bosea thiooxidans Calcium Phosphates Capsicum Cells Chlorophyll Cold Shock Stress Cold Temperature Crop, Avian Dietary Fiber DNA Replication Droughts Drought Tolerance Embryophyta Environment, Controlled Farmers Fertilization Glutamic Acid Glutamine Glycine Growth Disorders Herbaspirillum Herbaspirillum huttiense Leucine Lolium Lycopersicon esculentum Lysine Maize Massilia niastensis Methionine Microbial Consortia Nitrates Nitrites Nitrogen Novosphingobium rosa Paenibacillus Paenibacillus amylolyticus Pantoea agglomerans Pantoea vagans Phenotype Phosphates Photosynthesis Plant Development Plant Embryos Plant Leaves Plant Proteins Plant Roots Plants Polaromonas ginsengisoli Pseudoduganella violaceinigra Pseudomonas Pseudomonas fluorescens Rahnella Rahnella aquatilis Retention (Psychology) Rhodococcus erythropolis Rosa Salt Stress Sodium Chloride Sodium Chloride, Dietary Stenotrophomonas chelatiphaga Stenotrophomonas maltophilia Stenotrophomonas rhizophila Stenotrophomonas terrae Sterility, Reproductive Strains Technique, Dilution Threonine Triticum aestivum Tryptophan Tyrosine Vegetables Zea mays
2
Dictyostelium Strains Suppress Soft Rot Pathogens
Not available on PMC !

Example 1

119 Dicty strains were screened for their ability to feed on Dickeya (Dd) or Pectobacterium (Pcc) at 10° C. This assay was performed by inoculating Dd or Pcc on a low nutrient medium (SM2 agar) that supports both bacterial and Dicty growth. Dicty spores from individual strains were then inoculated on top of the bacterial growth and incubated at 10° C. to mimic potato storage temperatures. Dicty strains that successfully fed on Dd or Pcc created visible clearings in the lawn of bacterial growth and ultimately produced sporangia (fruiting bodies) that rose from the agar surface. An example of the phenotype that was considered successful clearing of bacteria is shown in FIG. 3A. From this initial screen, 36 Dicty strains that were capable of feeding on both Dd and Pcc at 10° C. were identified (FIG. 1B).

Of the 36 strains capable of feeding on both Dd and Pcc, 34 came from the Group 4 Dictyostelids (FIG. 1). This group includes D. discoideum, D. giganteum, D. minutum, D. mucoroides, D. purpureum, and D. sphaerocephalum (72). The results indicate that this group is particularly enriched in Dd and Pcc-feeding strains.

A further experiment was performed to identify Dicty species capable of feeding on biofilms of Dd and Pcc. Microporous polycarbonate membranes (MPMs) are widely reported to support biofilm formation of numerous Enterobacteriaceae species (2, 63, 70, 71). It was determined if Dd and Pcc formed biofilms on MPMs and determined if Dicty strains were capable of feeding on these biofilms. Membranes were placed on top of SM2 agar to provide Dd and Pcc with nutrients for growth. Bacteria were then inoculated on the surface of the MPMs and growth was monitored over the course of 1 week by washing bacteria off the membranes and performing dilution plating for colony counting. Growth of both bacterial strains plateaued around 4 dpi (FIG. 2).

From these results, it was determined that the best time to collect inoculated MPMs for biofilm analysis was at 2 dpi. Scanning electron microscopy (SEM) is commonly used to confirm biofilm formation by detecting extracellular polymeric substance (EPS) that forms the biofilm matrix (2). Samples of Dd and Pcc after 2 days of growth on MPMs in the presence and absence of Dicty are analyzed using SEM.

19 Dicty strains identified as active were tested for their ability to feed on Dd and Pcc growing on MPMs. These experiments were performed by establishing Dd and Pcc growth on MPMs overlaid on SM2 agar at 37° C. for 24 hr. Dicty spores were then applied to the center of bacterial growth in a 5 uL drop containing 1000 spores. Bacteria and Dicty were incubated at 10° C. for 2 weeks before remaining bacteria were washed off and colonies were counted. Representative images of Dicty growing on Dd and Pcc on MPMs are shown in FIG. 3A.

No Dicty strains produced a statistically significant reduction in Dd viability compared to the non-treated control. However, treating Dd lawns with Cohen 36, Cohen 9, WS-15, WS-20, and WS-69 consistently reduced the number of viable bacteria by approximately 100,000-fold compared to the non-treated control (FIG. 3B). Cohen 9 was the only Dicty strain that produced a statistically significant reduction in viability of Pcc compared to the non-treated control (FIG. 3C). Other Dicty strains capable of reducing the number of viable Pcc by at least 100,000-fold were Cohen 35, Cohen 36, WS-647, and WS-69 (FIG. 3C).

It was observed that Dicty strains Cohen 9, Cohen 36, and WS-69 were capable of feeding on both Dd and Pcc when these bacteria were cultured on SM2 agar and MPMs (FIGS. 1 and 3). These strains were also particularly effective feeders as all three reduced the number of viable Dd and Pcc on MPMs at 10° C. by 100,000-fold compared to the non-treated control (FIGS. 3B and 3C).

To determine if these strains could suppress soft rot development on seed potato tubers, tubers were tab-inoculated with Dd or Pcc and treated with spores from each Dicty strain. Seed potatoes were surface-sterilized and punctured using a sterile screw to a depth of 1.5 mm. Overnight cultures of Dd and Pcc were suspended in 10 mM potassium phosphate buffer, diluted to an OD600 of approximately 0.003, and administered as a 5 μL drop into the wound. Next, 5 of a Dicty spore suspension (100,000 spores) was added to the wound. Inoculated seed potatoes were placed in a plastic container with moist paper towels and were misted with water twice a day to maintain a high humidity. After 3 days at room temperature, seed potatoes were sliced in half and the area of macerated tissue was quantified using ImageJ.

All three strains reduced the severity of soft rot caused by Dd and Pcc (FIG. 4). Cohen 36 was the most effective strain on both Dd and Pcc: reducing the area of tissue maceration by 60% and 35%, respectively (FIG. 4B). Treating seed potatoes with WS-69 reduced the area of tissue maceration by 50% and 30% for Dd and Pcc, respectively (FIG. 4B). Finally, Cohen 9 was the least effective, but still able to reduce tissue maceration caused by Dd and Pcc by 25% and 20%, respectively (FIG. 4B).

FIG. 7 shows that three Dicty isolates control Dd and Pcc in seed tubers (at 25° C.). Two sets of data from different weeks were normalized to the Dickeya or Pectobacterium only bacterial control. The average area of macerated potato tissue measured in mm2 was set as “1” or “100%”. The average of all the other treatments including Dicty were divided by bacteria only control and multiplied by 100 to obtain a percentage. Each set contained 5 tubers per treatment.

Dicty should be capable of sporulating at temperatures as cold as 10° C. on a potato surface if they are applied as a one-time pre-planting or post-harvest treatment. Sporulation was assessed by inoculating small potato discs (5×6 mm) with 10 μL of Dd or Pcc suspensions at an OD600 of 3×10−5 and Dicty spores at a concentration of 1×107 spores/mL. Potato discs were kept in a covered 96-well plate for two weeks at 10° C. followed by visual inspection for son using a dissecting microscope. Representative images of a strain producing many sori (WS-517) and a strain producing few sori (WS-69) are shown in FIG. 5. Of the 11 strains evaluated, only Cohen 9 and WS-20 were unable to sporulate in the presence of both pathogens (Table 1).

TABLE 1
Assessment of Dicty sporulation at 10° C. on potato
in the presence of Dd or Pcc. A (✓) indicates sori
have been observed while a ( [Figure (not displayed)]  ) means they have not.
Dicty strainDdPcc
Cohen 9✓[Figure (not displayed)]
Cohen 36✓✓
WS-69✓✓
WS-517✓✓
WS-588✓✓
WS-606✓✓
WS-15✓✓
WS-20[Figure (not displayed)] ✓
DC-7✓✓
DC-61✓✓
WS-116d✓✓

Example 2

This example describes the use of a high throughput screening assay to identify Dicty strains from Alaska (e.g., BAC10A, BAF6A, BAC3A, NW2, KB4A (ATCC® MYA-4262™) SO8B, SO3A, BAF9B, IC2A (ATCC® MYA-4259™), AK1A1 (ATCC® MYA-4272™) PBF4B (ATCC® MYA-4263), PBF8B, BSB1A, SO5B (ATCC® MYA-4249), PBF3C, PBF6B, NW2B, NW10B (ATCC® MYA-4271™), PBF9A, IC5A (ATCC® MYA-4256TH), ABC8A (ATCC® MYA-4260), NW16B, ABC10B, ABB6B (ATCC® MYA-4261), BA4A (ATCC® MYA-4252), AKK5A, AKK52C, HP4 (ATCC® MYA-4286), HP8 (ATCC® MYA-4284), or NW9A) that feed on Dd and Pcc at 10° C. on potatoes.

Results from 11 Dicty strains screened against Dd at 10° C. are presented in FIG. 6. Data was analyzed for significance using a one-way analysis of variance (ANOVA; alpha =0.05) with Tukey's honest significant difference (HSD) test to compare means between the treatments and the No Dicty control. A reduction in Dd proliferation when potato discs were treated with Dicty strains Cohen 9, Cohen 36, WS-15, Maryland 18a, BAF6A, NW2, and SO3A.

The Alaskan Dicty strains, and those identified in Example 1, are further tested against coinfections of Dd and Pcc. It is useful to identify Dicty strains that can suppress Dd and Pcc coinfections as these two pathogens have been isolated together from diseased potatoes (15). The ability of Dicty strains with different feeding preferences (Dd vs. Pcc) to complement each other when administered as a cotreatment is assayed.

US11864559B2. Therapeutic amoeba and uses thereof (2024-01-09). AmebaGone, LLC [US]. Inventors: Marcin Filutowicz [US], Ryan Kessens [US], Amy Jancewicz [US].
Full text: Click here
Patent 2024
A-A-1 antibiotic Agar Amoeba Bacteria Biofilms Buffers Coinfection Cold Temperature Combined Modality Therapy Dickeya Dictyosteliida Enterobacteriaceae Extracellular Polymeric Substance Matrix Extracellular Polymeric Substances High-Throughput Screening Assays Human Body Humidity Microscopy neuro-oncological ventral antigen 2, human Nutrients Pathogenicity Pectobacterium Phenotype Plant Tubers polycarbonate potassium phosphate Scanning Electron Microscopy Solanum tuberosum Sporangia Spores Sterility, Reproductive Strains Technique, Dilution Tissue, Membrane Tissues Wounds
3
Synthesis of Bifunctional Urethane Acrylate
Not available on PMC !

Example 1

In a four-necked flask of 1 liter equipped with a well-dried stirring blade and a thermometer, 372 g (3.20 mol) of HEA, 0.71 g of DBTDL (0.1% by mass with respect to the total mass of HEA and TMHDI), and 0.35 g of MEHQ (0.05% by mass of the total mass of HEA and TMHDI) were added and stirred until the mixture was homogeneous, and then the temperature was raised to 60° C. Subsequently, 337 g of TMHDI (1.60 mol) was added dropwise over 1 hour. As the internal temperature rose due to the reaction heat during the drop, the drop rate was controlled so that the temperature was below 80° C. After completion of the dropping of the total amount, the reaction was carried out for 10 hours while keeping the temperature at 80° C. At this time, the proceedings of reaction were tracked by HPLC analysis to confirm the endpoint of the reaction. The product was discharged from the reactor to obtain 680 g of a bifunctional urethane acrylate (UDA). The viscosity at 25° C. was 7,100 mPa·s.

US11866527B2. Photocurable composition, cured product, and dental product (2024-01-09). MITSUI CHEMICALS, INC. [JP]. Inventors: Toshikazu Sakamaki [JP], Mai Kimura [JP], Hiroki Murai [JP], Takaaki Hayashi [JP], Suguru Endo [JP].
Full text: Click here
Patent 2024
High-Performance Liquid Chromatographies Thermometers urethane acrylate Viscosity
4
Synthesis of Bifunctional Urethane Acrylate
Not available on PMC !

Example 3

In a four-necked flask of 1 liter equipped with a well-dried stirring blade and a thermometer, 444 g (2.00 mol) of M-600A, 0.63 g of DBTDL (0.10% by mass with respect to the total mass of M-600A and XDI), and 0.32 g of MEHQ (0.05% by mass of the total mass of M-600A and XDI) were added and stirred until the mixture was homogeneous, and then the temperature was raised to 60° C. Subsequently, 188 g of XDI (1.00 mol) was added dropwise over 1 hour. As the internal temperature rose due to the reaction heat during the drop, the drop rate was controlled so that the temperature was below 80° C. After completion of the dropping of the total amount, the reaction was carried out for 10 hours while keeping the temperature at 80° C. At this time, the proceedings of reaction were tracked by HPLC analysis to confirm the endpoint of the reaction. The product was discharged from the reactor to obtain 600 g of a bifunctional urethane acrylate (MMD-352). The viscosity at 65° C. was 6,210 mPa·s.

US11866527B2. Photocurable composition, cured product, and dental product (2024-01-09). MITSUI CHEMICALS, INC. [JP]. Inventors: Toshikazu Sakamaki [JP], Mai Kimura [JP], Hiroki Murai [JP], Takaaki Hayashi [JP], Suguru Endo [JP].
Full text: Click here
Patent 2024
High-Performance Liquid Chromatographies Thermometers urethane acrylate Viscosity
5
Enhancing CRISPR-mediated HDR Efficiency
Not available on PMC !

Example 7

A mouse model was used to test HDR efficiency of the modified DNA donor molecules in vivo. A 4.6 kb long traffic light reporter (TLR), was integrated, or knocked-in, into the ROSA-26 locus. HDR efficiency was determined using donors either as plasmid or end-modified double-stranded PCR products. Using donors with 2′-OMe-RNA-TEG 5′ end-modifications, it was found that 9 out of 15 (60%) founder animals had the insert compared to 1/11 (9%) using plasmid as a donor. This result indicates that end-modifications make the donor molecules more potent even in mammalian (in vivo) settings (FIG. 23). The injection mixtures for this experiment are as follows: plasmid donor experiment—Cas9 mRNA (50.0 ng/μl), Cas9 protein (50.0 ng/μl), sgRNA (20 ng/μl), plasmid DNA donor (10 ng/μl); end-modified donor experiment—Cas9 mRNA (50.0 ng/μl), Cas9 protein (50.0 ng/μl), sgRNA (20 ng/μl), modified DNA donor (1 ng/μl).

US11873487B2. Compositions and methods for improved gene editing (2024-01-16). UNIVERSITY OF MASSACHUSETTS [US]. Inventors: Craig Mello [US], Krishna Sumanth Ghanta [US].
Full text: Click here
Patent 2024
Animals CRISPR-Associated Protein 9 Donors Light Mammals Mus Plasmids RNA, Messenger Rosa Tissue Donors

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More about "Rosa"

The Rosa genus, part of the Rosaceae family, is a diverse and versatile group of flowering plants renowned for their captivating blooms and alluring fragrance.
These shrubs, often referred to as roses, have been widely cultivated worldwide for their ornamental value, with a vast array of species and cultivars offering a range of colors, forms, and growth habits.
Roses have long held cultural significance and are frequently used in landscaping, cut flower arrangements, and perfume production.
Researchers may leverage the Rosa genus in various applications, such as the study of plant biology, the development of novel horticultural traits, or the extraction of valuable compounds.
The MeSH term provides a concise overview of the Rosa genus and its potential research applications.
Relatedly, the C57BL/6 and C57BL/6J mouse strains, along with the B6.Cg-Gt(ROSA)26Sortm14(CAG-tdTomato)Hze/J and B6.Cg-Gt(ROSA)26Sortm9(CAG-tdTomato)Hze/J genetically modified strains, have been widely used in biomedical research, often in conjunction with the MATLAB computational software.
The Tamoxifen compound has also been extensively studied for its various applications.
By incorporating insights from these related topics, researchers can gain a more comprehensive understanding of the Rosa genus and its potential for various applications, from ornamental horticulture to biomedical research.
This knowledge can be leveraged to optimize research protocols and identify the best products and resources for their needs, as facilitated by the PubCompare.ai platform.