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We narrowed to 80 results for: lentiviral infection

Showing: 21 - 40 of 80 results

  1. A Look Back at One Year of Plasmid Sharing for COVID-19 Research

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    Blog Post
    ...pseudovirus, scientists can study the mechanisms of infection and how to interfere with it in a BSL-2 lab instead...found that cathepsins play a role in SARS-CoV infection and deposited plasmids for the expression of cathepsin...SARS-CoV-2 entry and become upregulated during viral infection.  Expression of SARS-CoV-2 viral proteins In the...19 pandemic, the Krogan lab at UCSF generated lentiviral vectors expressing SARS-CoV-2 open reading frames...all but one are cloned into pLVX vectors (for lentiviral packaging) and contain an IRES-Puro marker for...as critical host factors modulating SARS-CoV-2 infection. bioRxiv. https://doi.org/10.1101/2021.04.22.440848...(2020) Protocol and Reagents for Pseudotyping Lentiviral Particles with SARS-CoV-2 Spike Protein for Neutralization...

  2. The AAVantages of AAV in CRISPR Screens

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    Blog Post
    ... screens rely on lentiviral systems, a fact supported by the wide array of lentiviral-based CRISPR libraries...vector delivery methods. Lentivirus has limited packaging options, with most lentiviral vectors being packaged...large packaging capacity and stable expression of lentiviral vectors make them an attractive option. However...into mice. For in vivo screens, mice are directly infected with the CRISPR library. In vivo screens can become...serotype has its own tropism, or a preference to infect specific cell or tissue types. Using serotypes ...example, AAV8 for kidney cells — can help improve infection rates in your experiment. Figure 2: Visual... while both AAV and lentivirus have low immunogenicity, AAV still beats lentivirus. In mice, AAV is almost...

  3. Multiplex Genome Editing with CRISPR-Cpf1

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    Blog Post
    ...Sakuma et al   Multiplexed Lentiviral Expression Cassettes lentiviral transduction One vector expresses...expressing the Cpf1 multiplex system from a lentiviral vector.  Lentiviruses carry a (+) strand RNA copy of the... multiple expression platforms: transfection, lentiviral transduction, and AAV transduction.  In most ...Cpf1. This means Cpf1 can bind and cleave the lentiviral RNA, preventing packaging of the virus.  Reversing...and in vivo. For these experiments, cells were infected with a 1:1 ratio of two AAVs,  one expressing ...a crRNA array plus a GFP tag. Four weeks post-infection, there was strong expression of Cpf1 and GFP in... options for Cpf1 multiplexing: Transfection, lentivirus, and AAV Cpf1’s ability to process its own pre-crRNA...

  4. Delivery Methods for Generating iPSCs

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    Blog Post
    ... for clinical applications.     Lentiviral: Lentiviral vectors infect dividing and nondividing cells and... a cancer-like phenotype in infected cells. Like retroviruses, lentiviral delivery is not considered safe...Excisable Cre-lox lentiviral vectors: This approach is the same as using a standard lentiviral vector, but ...retroviruses. Retroviral vectors, such as pMXs or pMSCV, infect dividing cells, and have a higher efficiency of...available from Addgene. Another drawback is that lentiviral vectors insert in different genetic locations...could be affected by the location of insertion. Lentiviral insertion can also disrupt the expression of ... Viral Vectors Adenovirus: Adenoviral vectors infect dividing and nondividing cells and have an ~8kb...

  5. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...cycles. Back to Top F. Infecting Target Cells Lentiviral particles can efficiently infect a broad range of ...Producing Lentiviral Particles E.1 Recommended materials E.2 Protocol for producing lentiviral particles...concentration F.3 Protocol for lentiviral infection and selection G. Safety H. References H.1 Published articles...transfection, or can be converted into lentiviral particles for subsequent infection of a target cell line. Once ...Addgene recommends that you generate lentiviral particles and infect the target cells. Addition of puromycin...puromycin concentration.) F.3. Protocol for Lentiviral Infection and Selection Day 1: a. Plate target cells...plate. TIP: MOI (multiplicity of infection) refers to the number of infecting viral particles per cell. Addgene...

  6. Lentivirus Plasmids

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    Collection
    ...target cell upon infection. When lentivirus is used for research, it is the lentiviral genome that encodes...Blog: Tips for Titering Your Lentiviral Preps Lentiviral Pooled Libraries All Viral Plasmids Packaging Plasmids...researcher safety. Read our Lentiviral Guide for more information about: Lentiviral components 2nd versus 3rd...Glossary of commonly used lentiviral terms Popular Lentiviral Envelope and Packaging Plasmids One safety feature...Reiser Return to Top Popular Lentiviral Transfer Plasmids The lentiviral genome (i.e., the genetic material...generation lentiviral systems Viral production Frequently asked questions about lentivirus use Glossary...table lists lentiviral transfer plasmids that can be used with 2nd and 3rd generation lentivirus technologies...

  7. Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection

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    Blog Post
    ...This can be done with lentiviral and Adeno Associated Viruses (AAV). Lentiviral and AAV can be used to...releasing the newly formed viral particles that can infect other bacteria. In the lysogenic cycle, the phage... transduction. Transferred DNA once inside the infected bacterium can either exist as transient extrachromosomal...division. You can find all kinds of different lentiviral and AAV plasmids as well as ready-to-use viral...

  8. Viral Vectors 101: Types of viruses

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    Blog Post
    ...transduce (infect) a wide range of cell types with a high efficiency. However, gamma-retrovirus infection is ...maximum titer, a plus for production and infection. This virus can infect dividing and non-dividing cells and...-studied lentivirus is HIV, which many lentiviral tools have been derived from. Lentiviruses have slightly...gamma-retrovirus, coming in around 9 kb. Unlike gamma, lentivirus can infect both dividing and non-dividing cells. Lenti...complementary DNA copies of their genome upon infecting a host cell. This viral DNA (also called proviral...to dividing cells only, so is not suitable for infection of quiescent cells or infrequently cycling cells... small, single-stranded DNA parvovirus. AAV can infect humans and some other mammals but is not currently...

  9. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

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    Blog Post
    ...minigenomes are incapable of producing infectious virus (similar to lentiviral vectors) because they do not encode... its own RNA-dependent RNA polymerase into the infected cells. The EBOV polymerase replicates the viral...as viral mRNAs would be in the course of viral infection), producing firefly luciferase. Firefly luciferase...absence of some viral proteins (compared to viral infection). Therefore, the roles/functions of individual...Addgene.org Browse Our Viral Vector Plasmids Read Our Lentiviral Vector Guide Read Our AAV Guide  ...Department of Microbiology, National Emerging Infectious Disease Laboratories.  She is interested in genome...

  10. Production of Virus in Insect Versus Mammalian Cells

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    Blog Post
    ...-293T cells. If you titrate your viral preps, lentiviral titration requires an additional couple of days...baculovirus, a family of viruses that specifically infect insects. This is what makes insect cells a popular...require cloning specific AAV or lentiviral machinery into a baculovirus plasmid. Production of baculovirus...baculovirus construct, called a bacmid. Like AAV and lentiviral packaging plasmids, bacmids are readily available... round of virus, called P1 (or sometimes P0), infecting new Sf9 cells, and getting an amplified P2 virus...cells Viruses AAV and lentivirus Baculovirus; can produce AAV and lentivirus if needed # of plasmids...Overcoming the Challenges of Lentivirus Production Tips for Titering Your Lentivirus Preps More resources on...

  11. Viral Vectors 101: Viral Vector Elements

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    Blog Post
    ... cargo into the host genome. Lentiviral-specific components Lentiviruses have gone through their fair ...protein that can also be pseudotyped to alter infectivity. Next, the virus must assemble and have appropriate...for gamma-retroviruses. While not required for lentiviral production, they can be helpful for certain applications... applications and aid in ease of use. Our lentiviral guide outlines these components.     Fig....Resources on Addgene.org Addgene's most popular lentiviral plasmids Addgene's most popular retroviral plasmids...Parts of the AAV packaging plasmid Overview of Lentiviral Packaging Plasmids Adenoviral production and ...function. Lentivirus/gamma-retrovirus Lentiviruses are a class of retrovirus (so all lentiviruses are retroviruses...

  12. 5 Tips for Troubleshooting Viral Transductions

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    Blog Post
    ...successfully transfected and produced infectious virus, they may get infected with this virus, and therefore,...number of infectious particles in a given viral stock will give you a sense of the infectivity of your ...An estimated 320,000 viruses can infect mammals. Even more abundant are the Earth’s estimated 1031 bacteriophages...bacteriophages (viruses that infect bacteria), many of which are doing important work in our microbiomes...al., 1999). 5. Check your packaging cells for infection If your transductions aren’t working, you can ...microscope and should appear fluorescent if they were infected. While there may be some low-level fluorescent...if the virus does not have the proper tropism to infect the packaging cells. Another method of ensuring...

  14. Multiplexed Overexpression of Regulatory Factors (MORF) collection

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    Blog Post
    ...Blog Learn More about Lentivirus Figure out How to Get the Most from Your Lentiviral Transduction Additional... individually overexpressed and available for lentiviral packaging and delivery to target cells as a pooled... Fig. 1: Barcoded TF library was packaged in lentiviral particles and used to transduce human embryonic...the Zhang lab controlled their multiplicity of infection so that a single TF was delivered to each cell...

  15. CRISPR 101: Mammalian Expression Systems and Delivery Methods

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    Blog Post
    ...efficiency. Lentiviral transduction Cas9 and gRNA can be present in a single lentiviral transfer vector...plasmids provide the necessary components to make lentiviral particles. Stable, tunable expression of Cas9...Transient or stable expression of SaCas9 and/or gRNA. Infects dividing and non-dividing cells. AAV is least toxic...: Traffic Light Reporter System: Can evaluate lentiviral component delivery as well as genome repair by...Difficulty Very easy Very hard Viral delivery   Lentivirus Retrovirus AAV Cells Dividing or nondividing...

  16. Retrovirus Plasmids

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    Collection
    ...Resources: γ-Retrovirus Guide Biosafety Guide Lentiviral Plasmids AAV Plasmids Viral Vectors 101 eBook...relatively high amounts of DNA (up to ∼8 kb), and infection results in long-term transgene expression. Some...

  17. Top Requested AAV of 2017: pmSyn1-EBFP-CRE

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    Blog Post
    ... and requests are still pouring in! While our lentiviral service is going strong, the AAV service has ...partially control which cells are successfully infected by an AAV by altering your injection site and ...

  19. 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 ...

  20. Plasmids 101: Gateway Cloning

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    Blog Post
    ...recombination reactions that take place when lambda phage infects bacteria. In vivo, these recombination reactions...Gateway cloning to generate our desired constructs: lentiviral expression of the human KRAS gene in mammalian...level, and experimental purpose. For mammalian lentiviral expression, we could use a vector like pLenti...available at Addgene, including the Frew Lab Multiple Lentiviral Expression Systems (MuLE) Kit, the MultiSite ...
Showing: 21 - 40 of 80 results