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Pseudotyped Viruses

Pseudotyped viruses are chimeric viruses that incorporate foreign viral envelope proteins into their capsid, allowing them to infect target cells that the original virus cannot.
These modified viruses are widely used in research to study virus entry, tropism, and neutralization, as well as in gene therapy and vaccine development.
The PubCompare.ai platform leverages advanced AI techniques to identify and compare the most reliable and effective pseudotvped virus protocols from scientific literature, preprints, and patents, saving researchers time and ensuring the accuracy of their work.
With PubCompare.ai, users can quickly find the best protocols for their needs, optimized for reproducibility and accuracy.

Most cited protocols related to «Pseudotyped Viruses»

Neutralizing Antibody Titer Calculation

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Publication 2013

Antibodies, Neutralizing Cells Diet, Formula DNA Replication Genes, Reporter HIV Seropositivity Luciferases Plasma Pseudotyped Viruses Psychological Inhibition Vaccines Virus

Pseudovirus Production and Titration

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Publication 2013

Biological Assay Cells Cloning Vectors Culture Media DEAE-Dextran Epiphyseal Cartilage FuGene Genes, Reporter Giant Cells HEK293 Cells HIV-1 Luminescent Measurements Microscopy Plasmids Polypropylenes Promega Pseudotyped Viruses Sterility, Reproductive Technique, Dilution Transfection Vertebral Column Virus

Monocyte-Derived Dendritic Cell Infection

Monocytes were isolated and incubated with GM-CSF and IL-4 to induce dendritic cell differentiation. Pseudotyped viruses and virus-like particles were produced by transient transfection of 293FT cells using TransIT-293 (Mirus). Infections were performed by incubating 10⁵ MDDCs in 96 well U bottom plates in the presence of 8 µg/ml polybrene. Cell surface staining of activation markers was performed 48h after infection. shRNA vectors carrying GFP were transduced into fresh monocytes together with SIVVLP(G) and dendritic cell differentiation was induced. More than 90% of cells were routinely transduced and cells were challenged at day 4 with HDVIRESRFP(G) or other control PAMPs.

Manel N., Hogstad B., Wang Y., Levy D.E., Unutmaz D, & Littman D.R. (2010). A cryptic sensor for HIV-1 activates antiviral innate immunity in dendritic cells. Nature, 467(7312), 214-217.

Publication 2010

Cells Cloning Vectors Dendrites Dendritic Cells Differentiations, Cell Granulocyte-Macrophage Colony-Stimulating Factor Infection Macular Edema, Cystoid Monocytes Pathogen-Associated Molecular Pattern Molecules Polybrene Pseudotyped Viruses Short Hairpin RNA Transfection Transients Virion

Monocyte-Derived Dendritic Cell Infection

Manel N., Hogstad B., Wang Y., Levy D.E., Unutmaz D, & Littman D.R. (2010). A cryptic sensor for HIV-1 activates antiviral innate immunity in dendritic cells. Nature, 467(7312), 214-217.

Publication 2010

Comprehensive Viral Neutralization Analysis

The neutralization panel data (IC₅₀ and IC₈₀ values for specific monoclonal antibodies and pseudotyped viruses) were collected from 49 published neutralization studies, mostly from tables in PDF format provided in the supplemental materials. The viral data was collected from Los Alamos HIV Database and in some cases personal communication with the authors, and required careful consideration and systematization to resolve sequence name ambiguities between different laboratories. Antibody sequences were downloaded from GenBank, and links to the structures in Protein Data Bank are provided. Standard statistics are applied to tally and analyze neutralization results. Specifically, antibody associations with viral mutations are evaluated by Fisher's exact test, counting the number of antibody-resistant or antibody-sensitive viruses (above or below threshold of detection) and the presence or absence of specific amino acids or N-glycosylation motif in the viral sequence alignment position (12 (link)).

Yoon H., Macke J., West AP J.r., Foley B., Bjorkman P.J., Korber B, & Yusim K. (2015). CATNAP: a tool to compile, analyze and tally neutralizing antibody panels. Nucleic Acids Research, 43(Web Server issue), W213-W219.

Publication 2015

Amino Acids Antibodies Antibodies, Viral Monoclonal Antibodies Mutation Protein Glycosylation Pseudotyped Viruses Sequence Alignment Virus

Most recents protocols related to «Pseudotyped Viruses»

HIV Pseudovirus Neutralization Assay

HIV envs JRFL and NL4-3 in the pSVIII expression plasmid, as well as the env-deleted backbone plasmid pSG3ΔEnv, were obtained from the NIH HIV Reagent Program. CH505 env⁹⁷ (link) was synthesized (GenScript) and cloned into the pLenti-III plasmid. HIV pseudotyped virus was produced by transient co-transfection of HEK293T cells with env plasmid DNA and pSG3ΔEnv using 25 kDa PEI (Polysciences) as previously described.⁹⁸ (link) HIV-1 neutralization assays were performed as previously described,⁶⁷ (link) with slight modifications. In brief, TZM-bl target cells were seeded onto half-well 96-well white plates in 50 μL of growth medium (DMEM containing 10% FBS, 20 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin) at 1 × 10⁵ cells/mL and incubated overnight at 37°C. Pseudotyped viruses (24 μL) were co-incubated with a dilution series of B cell supernatant or monoclonal antibody (8 μL) at 37°C for 1 h. The maximum possible volume, 25% of total volume, was used for the mock samples. Subsequently, medium was aspirated from the TZM-bl cells and 25 μL of the virus/supernatant mixture was added. The plates were incubated at 37°C for 16 h, then 75 μL of fresh medium was added to minimize evaporation. Infectivity was determined 72 h post infection by lysing the cells, adding Bright-Glo Reagent (Promega), and measuring luciferase activity with a Synergy H1 plate reader (BioTek). HIV-1 strains used: NL4-3 (CXCR-4 tropic, clade B, tier 1A, T cell line adapted strain), JRFL (CCR5 tropic, clade B, tier 2 isolate), and CH505 (CCR5 tropic, clade C, tier 2 transmitted founder primary isolate).

Vamva E., Ozog S., Leaman D.P., Yu-Hong Cheng R., Irons N.J., Ott A., Stoffers C., Khan I., Goebrecht G.K., Gardner M.R., Farzan M., Rawlings D.J., Zwick M.B., James R.G, & Torbett B.E. (2023). A lentiviral vector B cell gene therapy platform for the delivery of the anti-HIV-1 eCD4-Ig-knob-in-hole-reversed immunoadhesin. Molecular Therapy. Methods & Clinical Development, 28, 366-384.

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Publication 2023

B-Lymphocytes Biological Assay CCR5 protein, human Cell Lines Cells CXCR4 protein, human Glutamine HIV-1 Infection Luciferases Monoclonal Antibodies Penicillins Plasmids Promega Pseudotyped Viruses Strains Streptomycin T-Lymphocyte Technique, Dilution Transfection Transients Vertebral Column Virus

Neutralizing Antibody Quantification via Pseudotyped SARS-CoV-2 Virus

The NAb titres were quantified by neutralization assays based on S-pseudotyped vesicular stomatitis virus (VSV) according to a previously described method [22 (link)]. In brief, 293 T cells were transfected with plasmids expressing the S protein of SARS-CoV-2 Wuhan, Beta, Delta, or Omicron BA.1 strain. Meanwhile, the cells were infected with VSV of which the G gene was replaced with the firefly luciferase (Fluc) reporter gene. Thus, the S protein was incorporated onto the surface of the defective VSV. The pseudotyped reporter viruses could mimic the attachment and entry of authentic SARS-CoV-2. Serum samples were serially diluted 3-fold from 1:30–1:7290 with Dulbecco’s modified eagle’s medium (DMEM) supplemented with 1% penicillin–streptomycin (PS), 25 mM HEPES, and 10% fetal bovine serum (FBS). The diluted serum samples were added with pseudotyped viruses at 650 TCID50 per well and incubated at 37 °C for 1 h. The mixtures were then added into Huh-7 cells cultured in 96-well plates at 2 × 10⁴ cells per well, followed by incubation in 5% CO₂ at 37˚C for 24 h. Finally, the luminescence was measured by Bright-GloTM luciferase detection reagent (Promega). The infection inhibition rates at each dilution were calculated according to the relative light unit (RLU) values as follows: inhibition rate = [1 – (average RLU of sample – average RLU of virus control) / (average RLU of virus control – average RLU of cell control)] × 100%. Based on the inhibition rate, the half maximal effective concentration (EC50) was calculated by Reed-Muench method.

Li P., Wang Q., He Y., Yang C., Zhang Z., Liu Z., Liu B., Yin L., Cui Y., Hu P., Liu Y., Zheng P., Wang W., Qu L., Sun C., Guan S., Feng L, & Chen L. (2023). Booster vaccination is required to elicit and maintain COVID-19 vaccine-induced immunity in SIV-infected macaques. Emerging Microbes & Infections, 12(1), e2136538.

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Publication 2023

Biological Assay Cells Eagle Fetal Bovine Serum Genes Genes, Reporter HEK293 Cells HEPES Infection Light Luciferases Luciferases, Firefly Luminescence Penicillins Plasmids Promega Pseudotyped Viruses Psychological Inhibition SARS-CoV-2 Serum spike protein, SARS-CoV-2 Strains Streptomycin Technique, Dilution Vesicular stomatitis Indiana virus Virus

Pseudotyped SARS-CoV-2 Neutralization Assay

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Publication 2023

Antibodies Antibodies, Viral Biological Assay Cells Culture Media DNA Replication Gene Expression Genes, Reporter HEK293 Cells Leukemia Luciferases Luciferases, Firefly Luminescence Monoclonal Antibodies Mus prisma Promega Pseudotyped Viruses SARS-CoV-2 SARS-CoV-2 B.1.351 variant SARS-CoV-2 B.1.617.1 variant Senile Plaques spike protein, SARS-CoV-2 Technique, Dilution Tissues Titrimetry Virion Virus Virus Diseases

SARS-CoV-2 Spike Pseudovirus Entry Inhibition Assay

HEK293T cells were transfected with 0.7 μg of different Spike plasmids along with 1.4 μg of HIV-1-NL4.3-inGluc at a 1:2 ratio using polyethylenimine transfection (42 (link)). Then, 48 to 72 h posttransfection, the culture supernatants were harvested. After removing the cell debris by spinning down at 3,000 × g for 10 min, the viruses were aliquoted and stored at −80°C.
Pseudovirions were transduced into various cell lines; 6 h postransduction, the media were changed. Gaussia luciferase activity was measured 48 to 96 h after infection to determine the relative infectivity or entry efficiency of the indicated viruses.
For the inhibition assay using pseudotyped viruses, HEK293T-ACE2-TMPRSS2 cells were pretreated with the indicated concentrations of EST/E-64D (E64d) (Sigma, 330005) or camostat mesylate (camostat) (Sigma, SML0057) for 1 h, followed by transduction with pseudovirions of interest in the presence of the same concentrations of the drugs. After changing the media at 6 h postinfection (hpi), the luciferase activity was measured at 48 and 72 hpi.
For the inhibition assay using infectious viruses, Vero-ACE2-TMPRSS2 cells were pretreated with the indicated concentrations of EST/E-64D (E64d) (Sigma, 330005) or camostat mesylate (camostat) (Sigma, SML0057) for 1 h, followed by infection with infectious viruses in the presence of the same concentrations of the inhibitors for 24 h. Subsequently, the cells were fixed with 3.7% formaldehyde for 1 h and analyzed by a Life Technologies Attune NxT flow cytometer.

Qu P., Evans J.P., Kurhade C., Zeng C., Zheng Y.M., Xu K., Shi P.Y., Xie X, & Liu S.L. (2023). Determinants and Mechanisms of the Low Fusogenicity and High Dependence on Endosomal Entry of Omicron Subvariants. mBio, 14(1), e03176-22.

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Publication 2023

ACE2 protein, human Biological Assay camostat camostat mesylate Cell Lines Cells Formaldehyde HIV-1 Infection inhibitors Luciferases Pharmaceutical Preparations Plasmids Polyethyleneimine Pseudotyped Viruses Psychological Inhibition TMPRSS2 protein, human Transfection Vero Cells Virus Virus Diseases Virus Internalization

SARS-CoV-2 Pseudovirus Neutralization Assay

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Publication 2023

ACE2 protein, human Biological Assay Cells Centrifugation Culture Media Eagle Edetic Acid Fetal Bovine Serum Gene Expression Genes HEK293 Cells HIV-1 Homo sapiens Infection Light Luciferases Luciferases, Firefly Luminescence Mutagenesis, Site-Directed Plasma Plasmids PRSS1 protein, human Pseudotyped Viruses Puromycin SARS-CoV-2 Serum Technique, Dilution Transfection Vaccines Vertebral Column Virus

Top products related to «Pseudotyped Viruses»

Lipofectamine 2000 by Thermo Fisher Scientific

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Lipofectamine 2000 is a cationic lipid-based transfection reagent designed for efficient and reliable delivery of nucleic acids, such as plasmid DNA and small interfering RNA (siRNA), into a wide range of eukaryotic cell types. It facilitates the formation of complexes between the nucleic acid and the lipid components, which can then be introduced into cells to enable gene expression or gene silencing studies.

Bright glo by Promega

Sourced in United States

Bright-Glo is a luminescent reagent designed for the detection and quantification of adenosine triphosphate (ATP) in biological samples. It is a sensitive and reliable tool for measuring cellular ATP levels, which can be used to assess cell viability, cytotoxicity, and other cellular processes.

Britelite plus reagent by PerkinElmer

Sourced in United States

BriteLite Plus Reagent is a laboratory product manufactured by PerkinElmer. It is designed to facilitate luminescent reactions for various analytical applications. The core function of the reagent is to generate a luminescent signal, which can be detected and measured using appropriate laboratory equipment.

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Lipofectamine 3000 is a transfection reagent used for the efficient delivery of nucleic acids, such as plasmid DNA, siRNA, and mRNA, into a variety of mammalian cell types. It facilitates the entry of these molecules into the cells, enabling their expression or silencing.

Luciferase assay system by Promega

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The Luciferase Assay System is a laboratory tool designed to measure the activity of the luciferase enzyme. Luciferase is an enzyme that catalyzes a bioluminescent reaction, producing light. The Luciferase Assay System provides the necessary reagents to quantify the level of luciferase activity in samples, enabling researchers to study biological processes and gene expression.

Luciferase assay substrate by Promega

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The Luciferase assay substrate is a reagent used to measure the activity of luciferase, an enzyme that catalyzes a bioluminescent reaction. The substrate provides the necessary components to enable the luciferase reaction, allowing for the quantification of luciferase expression or reporter gene activity in biological samples.

Glomax 96 microplate luminometer by Promega

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Dulbecco modified eagle medium (dmem) by Thermo Fisher Scientific

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DMEM (Dulbecco's Modified Eagle's Medium) is a cell culture medium formulated to support the growth and maintenance of a variety of cell types, including mammalian cells. It provides essential nutrients, amino acids, vitamins, and other components necessary for cell proliferation and survival in an in vitro environment.

Luciferase lysis buffer by Promega

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Luciferase lysis buffer is a solution used to extract and solubilize luciferase enzymes from cells or tissue samples. It is designed to facilitate the measurement of luciferase activity, which is commonly used as a reporter in various biological assays.

Fetal bovine serum (fbs) by Thermo Fisher Scientific

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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.

What are the different types or variations of Pseudotyped Viruses?

Pseudotyped viruses can incorporate a wide variety of foreign viral envelope proteins, including those from retroviruses (e.g. HIV, MLV), filoviruses (e.g. Ebola), rhabdoviruses (e.g. VSV), and more. This allows them to be engineered to target specific cell types or entry pathways that the original virus cannot. The choice of envelope protein is a key parameter that determines the tropism and properties of the pseudotyped virus.

What are some common applications of Pseudotyped Viruses in research?

Pseudotyped viruses are widely used to study virus entry, tropism, and neutralization. They enable researchers to safely investigate the virus-host cell interactions without the risks associated with the original virus. Additionally, pseudotyped viruses are utilized in gene therapy and vaccine development, as the foreign envelope proteins can be leveraged to deliver genetic payloads or present antigens to the immune system.

What are some common challenges or considerations when working with Pseudotyped Viruses?

One key challenge is ensuring the pseudotyped virus maintains the desired properties and functionality of the foreign envelope protein. Factors like the choice of packaging cell line, purification methods, and storage conditions can all impact the virus' characteristics. Additionally, the pseudotyping process can sometimes alter the virus' tropism or entry efficiency compared to the original. Careful optimization and validation are crucial when using pseudotyped viruses for specific applications.

How can PubCompare.ai help researchers working with Pseudotyped Viruses?

PubCompare.ai's AI-driven platform allows researchers to more efficiently screen the literature and identify the most effective protocols for their Pseudotyped Virus needs. The platform's advanced comparisons can highlight key differences in protocol effectiveness, such as tropism, entry efficiency, and reproducibility. This enables researchers to choose the best option to ensure the accuracy and reliability of their work. PubCompare.ai can save researchers time and effort by guiding them to the optimal Pseudotyped Virus protocols for their specific research goals.

What are some tips for optimizing the use of Pseudotyped Viruses in research?

When working with Psudotyped Viruses, it's important to carefully validate the properties of the modified virus, including its tropism, entry efficiency, and any potential alterations compared to the original virus. Researchers should also consider factors like the choice of packaging cell line, purification methods, and storage conditions, as these can impact the virus' functionality. Additionally, running parallel controls with the unmodified parental virus can help isolate the effects of the pseudotyping. With the right optimization and validation, Pseudotyped Viruses can be a powerful tool for a variety of research applications.

More about "Pseudotyped Viruses"

Pseudotyped viruses are a fascinating area of research, with broad applications in virology, gene therapy, and vaccine development.
These chimeric viruses incorporate foreign viral envelope proteins into their capsid, allowing them to infect target cells that the original virus cannot.
This provides researchers with a powerful tool to study virus entry, tropism, and neutralization.
One of the key challenges in working with pseudotyped viruses is identifying the most reliable and effective protocols.
This is where the PubCompare.ai platform comes in.
Leveraging advanced AI techniques, PubCompare.ai can quickly identify and compare the best pseudotyped virus protocols from scientific literature, preprints, and patents, saving researchers valuable time and ensuring the accuracy of their work.
Researchers can use PubCompare.ai to find the optimal protocols for their needs, optimized for reproducibility and accuracy.
This can be especially helpful when using common laboratory reagents like Lipofectamine 2000, Bright-Glo, BriteLite Plus Reagent, Lipofectamine 3000, Luciferase Assay System, Luciferase assay substrate, and the GloMax 96 Microplate Luminometer, all of which are commonly used in pseudotyped virus research.
By incorporating foreign viral envelope proteins, pseudotyped viruses can be engineered to target specific cell types, making them a powerful tool for gene therapy applications.
They can also be used in vaccine development, as the modified viruses can be used to deliver antigens and elicit an immune response.
Overall, the world of pseudotyped viruses is a rapidly evolving field, with countless possibilities for advancing our understanding of viral entry, tropism, and neutralization.
With the help of platforms like PubCompare.ai, researchers can stay at the forefront of this exciting area of research, working with the most reliable and effective protocols to ensure the accuracy and reproducibility of their work.