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We narrowed to 79 results for: lentiviral infection

Showing: 41 - 79 of 79 results

  1. Fluorescence Titering Assay

    Type
    Protocol
    ...the titer of your lentiviral vector allows you to control the multiplicity of infection (MOI) in downstream...rapidly thaw the lentiviral aliquot on ice prior to use. Prepare dilutions of the lentivirus into DMEM complete...Protocol that can be used to titer lentiviral preparations carrying a fluorescence marker. Protocols...Introduction This protocol can be used to titer lentiviral preparations that carry a fluorescence marker... cell lines. It is not recommended to subject lentiviral supernatants to multiple freeze-thaw cycles. ...polyethersulfone filter to remove cells and debris. Lentiviral titer can decrease during cycles of freeze-thaw..., this protocol was developed using low titer lentiviral vectors (10 5 TU/mL). If you anticipate that ...

  2. Hot Plasmids: Spring 2025

    Type
    Blog Post
    ...psPAX2 (Addgene #12260). HeLa cells were infected with 96 h lentiviral supernatant and selected with puromycin...tailored deactivation rates Addgene's own empty lentiviral backbones New recombinant antibodies for Glypicans...very own set of empty, HA-, and EGFP-expressing lentiviral backbones designed right here at Addgene. Here's...launching later this year!  Find Addgene's own Lentiviral backbone plasmids here! Figure 2: Addgene...HA-EGFP Puro (Addgene #236081), then generated lentiviral particles in HEK293T cells with pMD2.G (Addgene...

  3. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...cells that have taken up the lentiviral transgene. Note that not all lentiviral vectors deliver antibiotic...efficiency. Infection efficiency will vary between cell lines. It is not recommended that lentiviral supernatants...Thaw the lentiviral aliquot on ice prior to use. Prepare a range of dilutions of the lentivirus in DMEM.... This is because lentiviral integration is random. Given the MOI of the lentivirus used, cells may also...an integrated lentiviral vector. Protocols...expressing a gene of interest from an integrated lentiviral vector. Unlike the short-term protein expression...transfection approaches, generating cell lines using lentiviral vectors enables long-term protein expression ...

  4. Colony Formation Titering Assay

    Type
    Protocol
    ...titer of your lentiviral preparation allows you to control the multiplicity of infection (MOI) in downstream...thaw the lentiviral aliquot on ice prior to use. Prepare 10-fold serial dilutions of the lentivirus into ...This protocol can be used to titer lentiviral preparations that confer antibiotic resistance. Protocols...between cell lines. It is not recommended that lentiviral supernatants be subjected to multiple freeze-...polyethersulfone filter to remove cells and debris. Lentiviral titer can decrease during cycles of freeze-thaw...transduced with the indicated serial dilutions of the lentiviral vector pRosetta . The following day, cells were...transduced with the indicated serial dilutions of the lentiviral vector pHAGE EF1α dCas9-KRAB . The following ...

  7. Viral Production at Addgene

    Type
    Blog Post
    ...Concentrated lentiviral preparations (“-LVC” catalog items) are generated from lentiviral preparations...being frozen. Specifically, lentiviral particles are collected from the lentiviral preparation by precipitation...for in our reported titers. Lentiviral vectors are titered using a lentiviral droplet digital PCR (ddPCR...Browse our AAV vector collection. Lentivirus Production Lentiviral preparations ("-LV” catalog items)...of the transfer plasmid.   Lentivirus resources Browse lentiviral preps available from Addgene. After... Feschenko, M. (2018). Determination of Lentiviral Infectious Titer by a Novel Droplet Digital PCR Method...through how we produce both our AAV preps and our lentiviral preps.   Viruses are generated using standard...

  8. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...the authors packaged it into lentivirus and used the resulting lentiviral library to generate a population...Science. 2013. PubMed PMID: 23328393 Wu lab human lentiviral CRISPR library     CRISPR pooled libraries enable...regulate the ability of WNV to induce cell death in infected cells (SEL1L, UBE2J1, EMC3, EMC2, DERL2, UBE2G2...Library. This complex second-generation CRISPR lentiviral library targets nearly all human protein-coding...from the CDC and cloned into a third generation lentiviral transfer vector. Each gene has been FLAG-tagged...apicomplexan organism Toxoplasma gondii to the infection of human fibroblasts. Toxoplasma mutants were ... genes were investigated for functions during infection of human cells, and one of them called “claudin-like...

  9. Viral Vectors 101: Virus Safety

    Type
    Blog Post
    ...continue the infection cycle once inside the host. While the virus can still enter (and thus infect) cells,...useful because they’re derived from viruses that can infect people, cells, and animals. Some of these viruses...how to keep the replication of virus and its infectivity in check. Engineering controls, safety systems...or anyone else in the lab or community would be infected by a lab-grown virus, it's important to understand...integrate into the host genome as part of the viral infection process (though it can integrate at chromosome...viruses grown in the lab are gamma retrovirus, lentivirus, adenovirus, and adeno-associated virus (AAV)...humans, earning this virus a BSL-2 classification. Lentivirus (a type of retrovirus) and gamma retrovirus require...

  10. Empty Backbones - Choosing Your Perfect Plasmid Backbone

    Type
    Collection
    ... - Tet-inducible lentiviral Gateway destination vector for gene expression Lentiviral backbones made by... Tet-inducible lentiviral shRNA expression pLKO.1 - TRC cloning vector - Lentiviral shRNA expression...Blasticidin Mammalian pLX304 - 3rd gen lentiviral vector lentiSAMv2 - Lentiviral sgRNA cloning backbone pLKO.1-... Thy1.1 - Lentiviral shRNA expression Gentamicin Varies pJL-TRBO - An improved agroinfection-compatible...C-terminal HA tag pInducer20 - Tet-inducible HA-tagged lentiviral vector for ORF expression pDest-565 - Gateway...for mammalian expression pLV-mCherry - 3rd gen lentiviral plasmid expressing mCherry pBad-EBFP2 - Bacterial...dedicated Adeno-associated Virus (AAV) Plasmids page Lentiviral Can transduce both dividing and nondividing ...

  11. Kazuhiro Oka Lentiviral Vectors

    Type
    Collection
    ...deposited a series of lentiviral and adenoviral transfer vectors with Addgene. Lentiviral Transfer Vectors ...For more information on lentiviral and adenoviral vectors read our Lentiviral Vector Guide and Adenoviral... in our Lentiviral Plasmids and Adenoviral Plasmids collections Addgene Blog: Your Lentiviral Plasmid ...vectors. Download the Lentivirus Transfer Vector Production Protocol Lentiviral Vector ID Purpose pCDH-EF1...Lentiviral Vectors deposited by Kazuhiro Oka from the Baylor College of Medicine Gene Vector Core Depositor... Viral Vectors Gene Vector Core Viral Vectors Lentiviral Transfer Vectors Adenoviral Transfer Vectors ...Vectors Within this set of lentiviral transfer vectors you can find reporter plasmids to test whether or not...

  12. Top Requested Lentivirus and AAV of 2016

    Type
    Blog Post
    ...lentiCas9-Blast Most of Addgene’s lentiviral inventory is based on CRISPR tools, though we do provide a...requested lentivirus and AAV of 2016. The top viruses of 2016 were (drumroll please)... Top Lentivirus: lentiCas9... Cas9-expressing lentiviruses will no longer be carried over to the gRNA lentiviruses. This means that...lentiCas9-blast lentivirus from Addgene, you could use it in conjunction with gRNA lentiviruses easily made...Viral Service and began delivering ready-to-use lentivirus and adeno-associated virus (AAV) to scientists... 2016, we expanded our viral inventory to 25 lentiviruses and 25 AAVs. These viruses have been distributed... as we like to sayat Addgene, productivity is infectious. Curious which viruses researchers have found...

  13. Viral Vectors 101: What is a Virus (and a Viral Vector)?

    Type
    Blog Post
    ...survive by hijacking the machinery of the cells they infect. Once they enter a cell, they treat their viral...class of virus. From one, come many After a virion infects a cell, the single particle creates many copies...of lysing, but they do generally still kill the infected cell.) This is in contrast to bacteria and eukaryotes...tools… if one isn’t too bothered by the risk of infection, that is. Thankfully, through the use of engineering...the virus that eliminate or reduce the risk of infection — researchers were able to create far safer viral...after the viruses they’re derived from: AAVs, lentivirus, gamma retrovirus, and adenovirus. And now that...

  14. Using Addgene's New Viral Service for Your Research

    Type
    Blog Post
    ... Service Suggestion Form. Lentivirus As some of the most popular lentiviral vectors in the repository ...collection. We’ll provide a brief rundown of what lentiviral and AAV preps are available below, but you can... tools, we’re starting the Viral Service with lentiviral preps for the delivery of Cas9 endonuclease, ...with viruses including advice on how to titer lentiviral preps, troubleshoot viral transductions, and ...Virus in Your Research: A Primer for Beginners Lentiviral Vector Uses and Overview AAV: A Versatile Tool... of our new Viral Service, we’re distributing lentivirus (with many CRISPR tools included among the preps...we’ve got viruses to help you out. Of course, lentiviruses are used in experiments other than just those...

  15. Getting Started with Behavioral Neuroscience: Standardizing Experiments and Using Viral Vectors

    Type
    Blog Post
    ...brain region of interest. AAV or lentivirus?  Both AAV and lentiviral vectors are used for intracerebral...genomes per milliliter) and lentiviral vectors around 108 IU/ml (infectious units per ml)  in order to ...wider diffusion is required. On the other hand, lentiviral vectors are superior packaging vectors for larger...astrocytes, and AAV4 targets mostly astrocytes. Lentiviral vectors are pseudotyped in most cases with VSV-G...injection. Find more details about choosing AAV or lentiviral vectors in our previous blog posts.  Viral vector...injections are: Vector titer: The concentration of infectious viral particles must be high enough to maximize...

  16. Viral Vectors 101: An Introduction to AAV

    Type
    Blog Post
    ...vector; these capsids allow the virus to infect (or not infect) different cell types with some specificity...you can coinfect with multiple viruses to express genes or gene sets over 4.7 kb. Co-infection rates for...respectively, cell-specific gene expression and viral infection. Cre or Flp dependent switches control gene expression...neuro-permissive (Issa, 2023). Time to expression AAVs have to infect cells, hijack host cell machinery, and synthesize...essential receptor for adeno-associated virus infection."Nature 4:520(7588):108-12 (2016). PubMed PMID...? Check out Our Primer for Beginners Read Our Lentivirus FAQ Learn about Viral Vector Elements Resources...

  17. Adeno Associated Virus (AAV) for Cell and Gene Therapy

    Type
    Blog Post
    ... and requires co-infection of helper viruses like adenovirus for productive infection. AAV vectors are...innate immunity. AAV vectors are also capable of infecting a wide range of cell types including but not limited...may therefore already have ways to prevent AAV infection. For example, prevalence of neutralizing antibodies...the immune system and for enhanced abilities to infect target tissues. Improved specificity through control...whose work has focused on gene therapy for viral infections. She is particularly passionate about HIV cure...technologies have been investigated including lentiviruses, adenovirus, and inorganic delivery tools such...

  18. How Dry Ice Affects Viral Vector Sample pH & How to Avoid It

    Type
    Blog Post
    ...study tested AAV infectivity after a range of pH exposures. They showed that AAV infectivity was not altered...alter the infectivity of the virus. However, there was a substantial reduction in infectivity after 3 weeks...different types of tubes: the ones we use to store lentivirus and the ones we use to store AAV. Our results...effect on the pH of the samples stored in our lentivirus tubes (Fig. 1). However, once thawed, there was... 1st Time CRISPR User) Tips for Titering Your Lentivirus Preps Additional Resources on Addgene.org Check... lentivirus...

  19. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    ... has limited tropism in the retina, primarily infecting cells of the retinal pigmented epithelium (RPE...this gene can be targeted to RPE cells using a lentiviral vector and a clinical trial is ongoing. Moving...other viral vectors such as adenovirus (AdV) and lentivirus (LV) are known to transduce retinal cells, AAV...

  20. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    ...screen. We start by infecting millions of embryonic stem cells (ESCs) with a lentiviral vector, an attenuated...retrovirus in the same family as HIV. By design, our lentivirii are genetically programmed to CRISPR out the ...cells. Remove this pillar and the ESC dies. Order Lentivirus from Addgene If you’re interested in designing...

  21. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    ...transduced with Cas9 expressing lentivirus at the indicated multiplicity of infection (MOI). Anti-vinculin Western...performing and analyzing a gene edit using Addgene’s lentiviral CRISPR tools. Schematic overview of CRISPR...stably expressing Cas9 via transduction with Cas9 lentiviral vector, lentiCas9-Blast (52962-LV, Figure 1)....Lines with Lentivirus Isolating a Monoclonal Cell Population by Limiting Dilution Lentivirus Production...

  22. Viral Vectors 101: Viral Applications

    Type
    Blog Post
    ...applications Viruses work by infecting a host cell (the target of viral infection) and delivering a genetic...high infection frequency and the viral titer (the number of viral particles capable of infecting a host...is more than one type of virus to choose from (lentivirus, retrovirus, AAV, etc.). When you’ve decided ...

  23. Important Considerations When Using AAVs

    Type
    Blog Post
    ... possible elicited immune response from viral infection. Animal safety is a top priority in our lab so...which determines which tissues and cells an AAV can infect) can influence an AAV’s value greatly. Applying...largely due the ability of different serotypes to infect different cell types. In the CNS, it has been shown...), which is a possible mechanism for the AAV to infect other cell types not within the vicinity of the...virus serotypes: impact of larger genomes on infectivity and postentry steps." Journal of virology 79.15...facilities (Biosafety Level-1). Other possible tools, lentiviruses for example, are not approved for use in vivo...viruses, so they are resistant to alcohol-based disinfectants. A 10% bleach solution should be used to clean...

  24. Viral Vectors 101: Transductions

    Type
    Blog Post
    ...in functional virions. These viral particles can infect other cells, but since the majority of the viral...then be placed on whatever cells you would like to infect with the virus. Then the viral particles do the...perform the transduction with: either retrovirus, lentivirus, adenovirus, or AAV. Then, your desired DNA to... some viruses integrate into the host genome (lentivirus and retrovirus) while others are non-integrating...Addgene.org Generating Stable Cell Lines with Lentivirus Virus Handling and Storage Addgene Viral Services...

  25. Lentiviral Vector Guide

    Type
    Guide
    ...Vectors Lentiviral Vector Guide Lentiviral Vector Guide Jump To: Lentiviral Vectors Lentiviral Generations...Read our lentiviral guide to learn about lentiviral components, generations, lentiviral production, and...gamma-retroviruses can only infect mitotically active cell types. This means that lentiviruses can infect a greater variety... Things Lentivirus Lentiviral Vector Uses and Overview Overcoming the Challenges of Lentiviral Production...safety. For a summary of all lentiviral plasmid components, see the Lentiviral Plasmid Elements table . In... a wide range of infectivity for different cell types (known as tropism). Lentiviral vectors have gone...different generations. Lentiviral Generations First-generation First-generation lentiviral plasmids are largely...

  26. Lentivirus ddPCR Titration

    Type
    Protocol
    ...Bergelson S, Feschenko M. Determination of Lentiviral Infectious Titer by a Novel Droplet Digital PCR Method...be close to zero (A01). In this protocol, the lentiviral particles are serially diluted and used to transduce... of Lentivirus Vectors You may also like... Lentivirus Colony Formation Titration Assay Lentivirus Fluorescence...genome to calculate the infectious titer: $$T = {V*C*D\over v}$$ Where: T = Infectious titer, TU/mL V = Viruses...average infectious titer obtained from the appropriate dilutions to calculate the final infectious titer... PCR (ddPCR) for lentivirus vector titration. Protocols... Protocols ddPCR Titration of Lentivirus Vectors ddPCR Titration...

  27. Gamma-Retroviral Vector Guide

    Type
    Guide
    ...gamma-retroviruses can only infect mitotically active cell types. This means that lentiviruses can infect a greater variety...plasmids, as lentiviral vectors do. For more about lentiviruses, see Addgene's lentiviral vector guide...difference between lentiviruses and gamma-retroviruses is that lentiviruses are capable of infecting non-dividing... to the Retroviridae family. Gamma-retrovirus infection is associated with many serious illnesses. Common...gamma-retroviruses may not be efficiently packaged by lentiviral packaging systems, and vice versa. Gamma-retroviral...packaging gene env ; usually VSV-G due to wide infectivity VSV-G (vesicular stomatitis virus G protein) ...common envelope gene, due to a wide range of infectivity for different cell types (known as tropism). ...

  29. Using AAV for Neuronal Tracing

    Type
    Blog Post
    ...these methods are naturally occurring viruses that infect, persist, and migrate within neurons, and that ... moves retrogradely from a peripheral site of infection to the central nervous system where it replicates... added benefit of only labelling the subset of infected cells that also express Cre. Another advantage...vectors into a brain region, the AAV particles will infect neurons at different ratios, making these neurons...intracellular transport without actually causing infection or leading to transgene expression within the ...). AAV does not have a membrane envelope like lentivirus, but instead is contained by a capsid which is...

  30. Guide to Using Pooled Libraries

    Type
    Guide
    ... are infected at a very low multiplicity of infection (MOI) to increase the odds that an infected cell...cells of interest. With others, notably pooled lentiviral plasmid libraries, the plasmids must first be...adequately represented in the population, you’ll need to infect many more cells than the number of plasmids present...Examples of published selection mechanisms for lentiviral CRISPR libraries include: resistance to the anti-cancer...delivering as virus, make virus; this creates a pooled lentiviral CRISPR library. Apply pooled library to cells... do not survive the selection mechanism. You’ll infect two sets of cells and subject one set to selection...libraries! Genome-wide Screening Using CRISPR/Cas9 Lentiviral CRISPR Libraries Enable Genome-Scale, Knockout...

  31. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    ...categories (e.g., for a lentiviral vector to be used in mammalian cells, check Lentiviral and Mammalian Expression...be prone to recombination in bacteria) such as lentiviral, retroviral, and AAV plasmids, we recommend the...that I have disclosed all known hazardous and infectious properties of the submitted materials. As the...

  32. Immunology Research Plasmids and Resources

    Type
    Collection
    ...kill cells that have been infected by a foreign pathogen. They detect infected cells via receptors such...construct complex lentiviruses for modification of mammalian cells with a single viral infection from the lab...pathogens each day. Their ability to protect against infection depends on two systems that work in concert: the... the innate and adaptive immune systems. Upon infection, the immune system triggers several processes ...specific antigen, it finds and attacks pathogens or infected cells expressing that antigen. Antibodies produced...destruction of pathogens. Over the course of an infection, adaptive immune cells will expand and give rise...

  33. CRISPR Guide

    Type
    Collection
    ...CRISPR libraries What Are Pooled Lentiviral CRISPR Libraries? Pooled lentiviral CRISPR libraries are a heterogeneous...system, you’ll need to either co-infect with a Cas9-producing lentivirus or use a Cas9-expressing stable...genome-wide screens using CRISPR involves pooled lentiviral CRISPR libraries. Figure 8: Overview of genome-wide...heterogeneous population of lentiviral transfer vectors. In most cases, each plasmid contains an individual...Figure 8A), then cloned in a pooled format into lentiviral transfer vectors (Figure 8B). Libraries come ...library is suitable for your experiment. Although lentiviral libraries are the most popular method for CRISPR...our pooled library guide and our blog post on lentiviral pooled libraries . Browse Libraries: CRISPR Pooled...

  34. Viral Vectors 101: The AAV Retrograde Serotype

    Type
    Blog Post
    ...Berkeley engineered an AAV variant that efficiently infects neurons, directs high-level transgene expression...Addgene Blog Get Started with AAV Learn more about Lentivirus Find out More about Addgene's Viral Service ...Ready-to-use Virus for Your Research Visit Our Lentivirus Guide Page ...

  35. Viral Vectors 101: Biosensors

    Type
    Blog Post
    ...viral tropism—the type of cells the viral particles infect. This can direct your biosensor to specific tissues...ready to deliver your sensor, whether you choose lentivirus, gamma retrovirus, or AAV as your vehicle, we...

  36. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    ...different concentrations of virus, or multiplicity of infections (MOIs), to determine the optimal dose of AAV ...always make more AAV than I think I need. Unlike lentivirus, AAV requires a higher number of viral particles...Addgene.org Learn how Addgene produces AAV and lentivirus Check out Addgene's virus protocols Find AAV ...

  37. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    ...or determination of the median tissue culture infective dose (TCID50) (Zen et al., 2004).  To choose the...production process.  Helper-free production Unlike lentivirus, AAV does not possess the ability to replicate...are measured. A TCID50 measurement reports an infectious titer, meaning the concentration of viral particles...2010). It is recommended that you measure your infectious titer, since it will have more impact on your...Adeno-Associated Viral Vectors by Oxidant-Based Disinfectants. Human Gene Therapy, 32(13–14), Article 13–14...Bleu, T., Sommer, J. M., & Wright, J. F. (2004). Infectious titer assay for adeno-associated virus vectors... with sensitivity sufficient to detect single infectious events. Human Gene Therapy, 15(7), Article 7....

  38. Molecular Biology Reference

    Type
    Guide
    ...particles, such as lentiviral, retroviral, AAV, or adenoviral particles, that can infect your target cells...viral service offers select ready-made AAV and lentiviral particles. Visit our viral service page to learn...

  39. CRISPR Guide

    Type
    Guide
    ...CRISPR libraries What Are Pooled Lentiviral CRISPR Libraries? Pooled lentiviral CRISPR libraries are a heterogeneous...system, you’ll need to either co-infect with a Cas9-producing lentivirus or use a Cas9-expressing stable...genome-wide screens using CRISPR involves pooled lentiviral CRISPR libraries. Figure 8: Overview of genome-wide...heterogeneous population of lentiviral transfer vectors. In most cases, each plasmid contains an individual...Figure 8A), then cloned in a pooled format into lentiviral transfer vectors (Figure 8B). Libraries come ...library is suitable for your experiment. Although lentiviral libraries are the most popular method for CRISPR...our pooled library guide and our blog post on lentiviral pooled libraries . Browse Libraries: CRISPR Pooled...
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