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We narrowed to 75 results for: generation lentiviral packaging

Showing: 41 - 60 of 75 results

  1. Plan Your Experiment

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    Collection
    ...want to: Generate complete and permanent loss of gene expression or function ( knockout )? Generate a specific... Edit Generate a specific user-defined sequence change in a particular gene, such as generating a point...high efficiency Lentiviral transduction Cas9 and gRNA can be present in a single lentiviral transfer vector...positive cells Packaging and envelope plasmids provide the necessary components to make lentiviral particles...vector and used to generate AAV particles (for details, see our AAV Guide ) ∼4.5 kb packaging limit (only compatible...like never before. This section will provide a general framework to get you started using CRISPR in your... enrich positive cells, or selection marker to generate stable cell lines (for details, see Plasmids 101...

  2. Viral Vectors 101: AAV Serotypes and Tissue Tropism

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    Blog Post
    ... and infect specific cells. This protein shell packages the vector’s genetic material, and variations ...the viral particle sneaks its way into cells. Generally, AAV serotypes have a primary cellular receptor...and can be selected as a serotype for Addgene’s Packaged on Request service. Some AAV1 tools are available...and can be selected as a serotype for Addgene’s Packaged on Request service. Some AAV2 tools are available...central nervous system, including neurons, AAV5 is generally the best choice for targeting astroglia. It has...and can be selected as a serotype for Addgene’s Packaged on Request service. Some AAV5 tools are available...and can be selected as a serotype for Addgene’s Packaged on Request service. Some AAV8 tools are available...

  3. Adenovirus Plasmids

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    Collection
    ...then be transfected into mammalian packaging cell lines to generate adenovirus particles. Read our Adenoviral...Additional Resources: Adenovirus Guide Virus Biosafety Lentiviral Plasmids AAV Plasmids Viral Vectors 101 eBook... adenoviruses in research include their large packaging capacity (>8 kb), high titers, and high levels...terms Adenoviral Strains Bacterial strains for generating adenovirus. ID Strain Description PI 16398 BJ5183...cell types with almost 100% efficiency. Unlike lentiviruses or other retroviruses , adenoviruses do not ...

  4. Hot Plasmids: Spring 2025

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    Blog Post
    ...CMV HA-EGFP Puro (Addgene #236081), then generated lentiviral particles in HEK293T cells with pMD2.G (...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...taken them for a test drive! We validated viral packaging and strong expression after inserting EGFP and...and provide step-by-step protocols for viral generation and stable cell line creation (Figure 2). Looking...launching later this year!  Find Addgene's own Lentiviral backbone plasmids here! Figure 2: Addgene...Addgene #12260). HeLa cells were infected with 96 h lentiviral supernatant and selected with puromycin. Puromycin-resistant...

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

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    Blog Post
    ...COVID-19 pandemic, the Krogan lab at UCSF generated lentiviral vectors expressing SARS-CoV-2 open reading...but one are cloned into pLVX vectors (for lentiviral packaging) and contain an IRES-Puro marker for stable...from Alejandro Balazs’s lab includes those for generating pseudovirus and those encoding different spike...(2020) Protocol and Reagents for Pseudotyping Lentiviral Particles with SARS-CoV-2 Spike Protein for Neutralization...published a protocol and reagents for pseudotyping lentivirus with the SARS-CoV-2 spike protein. They deposited...

  6. Choosing Your Perfect Empty Backbone

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    Blog Post
    ...remember that you will need accessory packaging vectors to generate the virus. Bacterial vectors This system...vivo (try pcDNA3-EGFP). If you can get away with generating transiently transfected cells, you will not need...basic mammalian expression system and go for a lentiviral or a retroviral backbone. This is considered ...

  7. Viral Vectors 101: Types of viruses

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    Blog Post
    ...-studied lentivirus is HIV, which many lentiviral tools have been derived from. Lentiviruses have slightly...larger packaging capacities than gamma-retrovirus, coming in around 9 kb. Unlike gamma, lentivirus can infect... 1982). To add insult to injury, retroviruses generally have moderate to high immunogenicity, meaning ...interchangeable between the two systems, other, more general, viral components such as a heterologous envelope...adenovirus have been identified so far! Adenovirus generally has high immunogenicity, but some strains have... expression of adenovirus cargo is high, it’s generally not as long lasting as AAV (up to several weeks... fact: All viruses discussed in this blog are generally broadly infective, with one exception. Blood cells...

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

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    Blog Post
    ...analyses needed for packaging AAV. AAV production Overview: The first step in packaging AAV is transfecting...had never worked with AAV, so naturally I had to package some virus for my first experiment. It was a bit...transfecting HEK293 cells with AAV packaging plasmids. The cells need three different plasmids to produce AAV...intermediate into the single-stranded genome that's then packaged into AAV’s protein shell, or capsid; 2) the pHelper...plasmid which encodes a transgene of interest that’s packaged into the virus. Two to five days after transfection...plasmids contain ITRs which are required for proper packaging of AAV, but also form secondary structures that...spreadsheet similar to this one to plan my AAV packaging PEI transfections. It saves me time and helps ...

  9. 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...process, simply enter your preprint title or a general description of the deposit. Select your desired...millions. Pooled libraries must be verified by next-generation sequencing. Please see the help center article...that can be used under BSL1 conditions were generated in your lab are necessary for use of a plasmid...be prone to recombination in bacteria) such as lentiviral, retroviral, and AAV plasmids, we recommend the...with a unique Deposit ID. Addgene will send you a package containing instructions on how to prepare your ...

  10. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    ...sequence all the DNA packaged inside the viral particles (Figure 1). Briefly, packaged DNA is extracted from...more rigorous quality control, including next generation sequencing (NGS) of the viral genome to confirm... the helper plasmids or the cell genome can be packaged inside the vectors during production. These impurities... the virus itself. While these impurities are generally considered innocuous for research-grade vectors... to determine the identity and serotype of the packaged DNA and look for potential contaminants. The ...mentioned earlier, it is common to find DNA from the packaging cell genome, bacterial genomes, cloning vectors...the recombinase for transgene expression. Once packaged in AAV, this pre-recombined DNA could lead to ...

  11. CRISPR Library Amplification

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    Protocol
    ...as these sequences are not efficiently packaged into lentiviral particles. See below for options if the...amplification. This protocol is designed to be as general as possible but note that individual libraries ...our protocol on Bacterial Transformation . The generalized pooled library amplification protocol is expected... a diagnostic digest and high-throughput next generation sequencing (NGS). Select a restriction enzyme...gel ( see restriction digest protocol here ). Lentiviral plasmids can recombine between their LTRs resulting...

  12. Addgene: The First Twenty Years

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    Blog Post
    ...channel in 2013, which continue to grow today. “I generally refer people to Addgene for the basics of molecular...ready-to-use viral vectors from some of our most popular lentiviral and AAV vectors, as well as developing and sharing...expanding our viral vectors service to include AAV Packaged on Request! “I love Addgene. I think resources...

  13. Zhang Lab CRISPR Page

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    Collection
    ...#61425 - #61427 are intended for lentivirus production : 61425 : lentiviral vector encoding dCAS9-VP64 with...For cloning information, please view: Zhang Lab General Cloning Protocol 237.2 KB SpCas9 Plasmids The CRISPR... top GeCKO Screening Library GeCKO are pooled lentiviral CRISPR libraries (both human and mouse) for performing...with 2A Blast resistance marker 61426 : lentiviral vector encoding the MS2-P65-HSF1 activator helper complex...complex with a 2A Hygro resistance marker 61427 : lentiviral sgRNA cloning backbone with MS2 loops at tetraloop...SaCas9) that has been human codon-optimized, and packages both the SaCas9 gene and its single guide RNA ...of a single AAV vector ( AAV-KPL ) in the lung generated loss-of-function mutations in p53 and Lkb1 , as...

  14. CRISPR References and Information

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    Collection
    ... Information Addgene Resources CRISPR Software General Resources CRISPR Protocols Addgene CRISPR Resources...CRISPOR batch (Link opens in a new window) for lentiviral screens. sgRNA Scorer 2.0 (Link opens in a new...species. Developed by the Kate O'Connor-Giles Lab . General Resources CasPEDIA (Link opens in a new window)...Link opens in a new window) CRISPR/Cas9 reagent generation; gRNA design; HDR design; Deep sequencing Cas9...PDF, 255 KB Zhang sgRNA cloning lentiCRISPR v2 packaging plasmids: pVSVg , psPAX2 ; positive control: CMV-EGFP...vector system: lentiCas9-Blast and lentiGuide-Puro packaging plasmids: pVSVg , psPAX2 positive control: CMV-EGFP...

  15. Viral Vectors 101: Transductions

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    Blog Post
    ...viral packaging elements that will ensure the virus gets produced, and viral particles are packaged, when...Resources Resources on Addgene.org Generating Stable Cell Lines with Lentivirus Virus Handling and Storage Addgene...is then typically transfected along with viral packaging plasmids into a viral production cell line, usually...cell line, viral particles will be produced and packaged, resulting in functional virions. These viral ...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...

  16. Hot Plasmids: Winter 2025

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    Blog Post
    ...proportions from a vector backbone that allows lentiviral packaging as a single payload (Figure 4). ...applicable to dicot species. In addition, they generated offspring tomato plants that inherited the genomic...single-molecule fluorescence techniques. After generating a series of single- and double-tagged constructs...kinesins KIF5A and KIF5B as well as backbones for generating other SNAPf-, CLIPf-, and CLAP-tagged proteins...indicator ONE-GO biosensors kit Antibodies for neurodegeneration research   Optimized prime editing in dicots....01.028. New recombinant antibodies for neurodegeneration research   By Ashley Waldron We recently added... antibodies against important targets for neurodegeneration research: Anti-MAPT [AT8] and Anti-APP [6E10...

  17. Viral Vectors 101: An Introduction to AAV

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    Blog Post
    ... is a single stranded, Parvoviridae DNA virus, packaged in a non-enveloped icosahedral capsid, that can...the type of capsid (outer protein shell) used to package an rAAV vector; these capsids allow the virus to... over 4.7 kb. Co-infection rates for AAVs are generally high enough for this approach to work, although...design single rAAV vectors with multiple genes packaged in them and use elements such as IRES or 2A to...rescued from the vector plasmid and individually packaged into the AAV capsids. As a genome conversion in...neonatal mouse eyes at two different ages." Retinal Degenerative Diseases. Springer International Publishing,...? Check out Our Primer for Beginners Read Our Lentivirus FAQ Learn about Viral Vector Elements Resources...

  18. 27 Hot Plasmids from 2016

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    Blog Post
    ...the authors packaged it into lentivirus and used the resulting lentiviral library to generate a population... CRISPR Library. This complex second-generation CRISPR lentiviral library targets nearly all human protein-coding...obtained from the CDC and cloned into a third generation lentiviral transfer vector. Each gene has been FLAG-tagged... of dCas9-VPR limits one’s ability to package it into lentivirus and later create stable cell lines expressing...Science. 2013. PubMed PMID: 23328393 Wu lab human lentiviral CRISPR library     CRISPR pooled libraries enable... transfected with a Cas9-expressing plasmid to generate a population of mutant cells. The mutant cells...can be used to perform screens in human cells. General information regarding how to use a CRISPR pooled...

  19. Viral Vectors 101: Virus Safety

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    Blog Post
    ...? That’s okay! In this article we will discuss general viral safety considerations and best practices ...Gag structural protein on yet a third plasmid (packaging plasmid). Viral production then proceeds by transfection....   Figure 1: Separation of retroviral packaging components onto distinct plasmids for safety.  ... and the same holds true for types of viruses generated in the lab. The four most common types of viruses... lab and require additional safety measures.   General viral handling safety   Containment and personal...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...

  20. CRISPR Guide

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    ...using dual nickases to generate a knockout or gene edit; or using Cas9 to generate large genomic deletions...CRISPR libraries What Are Pooled Lentiviral CRISPR Libraries? Pooled lentiviral CRISPR libraries are a heterogeneous...’ll need to generate lentivirus containing the entire library (Figure 8D). This lentivirus will then be...nucleases (TALENs) required scientists to design and generate a new nuclease pair for every genomic target. ..., and precisely edit DNA and RNA. The ease of generating gRNAs makes CRISPR one of the most scalable genome...distal to the PAM often permit target cleavage. Generating a Knockout Using CRISPR Cas9 undergoes a second...The resulting DSB is then repaired by one of two general repair pathways: the efficient but error-prone ...
Showing: 41 - 60 of 75 results