Fbxl12 conditional knockout mice were generated with a targeting vector purchased from the Knockout Mouse Project (KOMP) repository (http:www.komp.org ). The vector was linearized and transfected into B6 embryonic stem (ES) cells. Transfected ES cells were cultured with media containing neomycin and resistant clones were screened for homologous recombination by PCR. Blastocyst injections resulted in several chimeric mice, three of which gave germline transmission. Germline Fbxl12fl/+ mice were crossed to ROSA26::FLPe mice (Jackson labs; Stock no. 003946) to delete the Neo gene. Offspring were then crossed to generate Fbxl12fl/fl mice. Fbxl1−/− mice51 (link) were provided by Dr.Liang Zhu (Albert Einstein College of Medicine). Cdkn1bfl/fl mice52 (link) were purchased from Jackson Laboratory (Stock no. 027328). Lck-Cre transgenic mice, AND TCR-transgenic mice, Rag2−/− mice and CD45.1 C57BL/6 mice were obtained from Taconic Biosciences. Animal experiments were approved by the Animal Care and Use Committee of the National Institute of Child Health and Human Development.
Rosa26 flpe mice
Manufactured by Jackson ImmunoResearch
The ROSA26::FLPe mice are a genetically modified mouse strain that expresses the FLPe recombinase enzyme from the endogenous ROSA26 locus. The FLPe enzyme can be used to induce site-specific recombination in mice, allowing for the temporal and spatial control of gene expression or deletion.
Automatically generated - may contain errors
3 protocols using rosa26 flpe mice
1
Generation of Fbxl12 Conditional Knockout Mice
2
Generation of Mettl3 Knockout Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Stem cell lines and mice deficient for Mettl3 were generated by targeted disruption of the endogenous Mettl3 locus via homologous recombination. The targeting strategy and construct Knockout Mouse Project repository [Mettl3:tm1a(KOMP)Wtsi] introduced loxP sites spanning the fourth exon that would result in an out-of-frame and truncated product upon deletion and introduced a LacZ reporter cassette driven by the endogenous Mettl3 promoter. Fifty micrograms of DNA of the targeting construct was linearized and electroporated into a V6.5 ESC line that was then subjected to selection with 300 µg/mL G418. After 10 d of selection, resistant clones were analyzed for correct targeting. Mettl3f/f floxed ESCs were injected to BDF1 host blastocyst and chimeric mice were generated. Chimeric male mice were mated with C57BL/6 females. F1 offspring were screened for germline transmission by agouti coat color and validation via PCR of LacZ transgene reporter. To remove neomycin and lacz cassette F1 offspring were mated with Rosa26-FlpE mice (Jackson Laboratory 003946) and offspring pups were validated for the removal of LacZ transgene. The mice were crossed to C57BL/6 for three generations before being used for any experiment.
3
Generation of Fbxl12 Conditional Knockout Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fbxl12 conditional knockout mice were generated with a targeting vector purchased from the Knockout Mouse Project (KOMP) repository (http:www.komp.org ). The vector was linearized and transfected into B6 embryonic stem (ES) cells. Transfected ES cells were cultured with media containing neomycin and resistant clones were screened for homologous recombination by PCR. Blastocyst injections resulted in several chimeric mice, three of which gave germline transmission. Germline Fbxl12fl/+ mice were crossed to ROSA26::FLPe mice (Jackson labs; Stock no. 003946) to delete the Neo gene. Offspring were then crossed to generate Fbxl12fl/fl mice. Fbxl1−/− mice51 (link) were provided by Dr.Liang Zhu (Albert Einstein College of Medicine). Cdkn1bfl/fl mice52 (link) were purchased from Jackson Laboratory (Stock no. 027328). Lck-Cre transgenic mice, AND TCR-transgenic mice, Rag2−/− mice and CD45.1 C57BL/6 mice were obtained from Taconic Biosciences. Animal experiments were approved by the Animal Care and Use Committee of the National Institute of Child Health and Human Development.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!