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Showing: 41 - 60 of 188 results
  1. Plasmids 101: The protein expression toolbox

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    Blog Post
    ...transiently induce expression? Check out Addgene's Lentiviral dCas9 SAM vector and Piggybac dCas9 SAM vector...promoters and modes of action Check out Addgene's Lentiviral Tet-on and AAVS1 targeted Tet-on transgene vector...Adapted from Nabet et al. Check out Addgene's Lentiviral dTAG cloning vector and SMASh degron tagging ...Addgene website Promoter Guide CRISPR Guide Lentiviral Guide  ...mechanism to integrate your choice cDNA. Both lentivirus and retrovirus can package cDNAs for genomic ...
  2. New Tool for Lineage Tracing: The ClonTracer Library

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    Blog Post
    ...individually label millions of mammalian cells through lentiviral infection and to monitor their abundance and ...optimize protocols for in vivo screening with lentiviral pooled shRNA libraries. Later, Bhang says, “When... treatment. The library consists of a pool of lentiviral vectors with roughly 73 million semi-random, ... Find the ClonTracer Library at Addgene Find Lentiviral Plasmids at Addgene Check out Our Cancer Resources...infection. When using the ClonTracer system, lentiviruses containing these barcodes infect the target ...
  3. Delivery Methods for Generating iPSCs

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    Blog Post
    ...considered safe for clinical applications.     LentiviralLentiviral vectors infect dividing and nondividing...Excisable Cre-lox lentiviral vectors: This approach is the same as using a standard lentiviral vector, but ...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 ...phenotype in infected cells. Like retroviruses, lentiviral delivery is not considered safe for clinical ...plasmids if you’re interested in Tet-inducible lentiviral expression of OSKM reprogramming factors: ...transgene reactivation, residual expression Lentivirus Integrating High Low Efficient and Stable Genome...
  4. 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...: 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...
  5. Top Requested Plasmid of 2017 - pMD2.G

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    ...2015 and 2016 were CRISPR plasmids), but so are lentiviral vectors, many of which are used to deliver Cas9...Transductions Getting the Most from Your Lentiviral Transduction Additional Resources on Addgene.org Check out... of 2017 is the lentivirus envelope plasmid pMD2.G from Didier Trono’s lab! Lentivirus envelope vector... producing our ready-to-use lentivirus preps. You can find our lentivirus production protocol on our website...titering your lentivirus in another blog post. If you need a quick primer on how lentiviruses work, check...promoter. Envelope proteins are a key component of lentiviruses. These proteins coat the virus and help determine...widely used and requested because it enables lentiviruses to infect many different types of cells. This...
  6. Introducing Addgene's New Search - Find Plasmids More Easily For Your Research!

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    Blog Post
    ...parentheses.  In this example I wanted to find a lentiviral plasmid to express ubiquitin under control of...300+ plasmids. Under ‘Vector Types’ I selected ‘Lentiviral’ which eliminates most of the plasmids designed...this filter will allow to refine by serotype or lentiviral prep Requests: Only see plasmids with more than... cell type Serotype: Choose the AAV capsid or lentiviral prep needed Tags: Identify a specific marker ...
  7. 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... while both AAV and lentivirus have low immunogenicity, AAV still beats lentivirus. In mice, AAV is almost...made at high titers equal to those achieved with lentivirus but offer an improved safety profile, customization...expression in many mouse tissues. Safety AAV and lentivirus have different safety profiles that often affect...experiments. AAV can be safely handled at BSL-1, while lentivirus requires a BSL-2 setup. This requires a less ...
  8. 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...
  9. Hot Plasmids Spring 2024

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    Blog Post
    ...within the lentiviral 3’ long terminal repeats (LTR). The 3’ LTR is duplicated during lentiviral integration...license. In this iteration, the size of the lentiviral insert was minimized to reduce recombination ...III transcript, the sgRNAs were encoded on the lentiviral minus strand in the 3’ LTR. These optimizations...
  10. Viral Vectors 101: Gamma-Retroviral Packaging Systems

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    Blog Post
    ... well as the viral LTRs, differ. As a result, lentiviral and γ-retroviral packaging plasmids are not interchangeable...using 293T cells This method is very similar to lentiviral packaging methods. The three components described...found within both the 5' and 3' LTRs of retro/lentiviral plasmids. U5; Unique 5'; region at the 5' end... cells a virus can infect. Note: While both lentiviruses and γ-retroviruses use the same gene products...How do engineered γ-retroviruses differ from lentiviruses? There are a few important differences between...between engineered γ-retroviruses and lentiviruses. First, these viruses were derived from different genomes...genomes (MoMLV and MSCV for γ-retrovirus; HIV for lentivirus). Additionally, γ-retroviruses can only transduce...
  11. Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

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    Blog Post
    ...conducted earlier by the Zhang lab for GeCKO, a lentiviral CRISPR library enabling genome-scale, knockout...that you review the steps for how to use pooled lentiviral CRISPR libraries and keep the following in mind... sequencing technology. These libraries are lentiviral in nature, so please ensure that you are equipped...equipped and authorized to make and use lentivirus. Also, note that due to the size of the backbone, transformations...
  12. Plasmids 101: Common Lab E. coli Strains

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    Blog Post
    ... NEB Stable   For cloning into and storage of lentiviral and retroviral vectors or cloning or repeated...JM109-derived. For cloning into and storage of lentiviral and retroviral vectors or cloning or repeated...Derived from HB101. For cloning into and storage of lentiviral and retroviral vectors or cloning or repeated...cells for cloning and storage of unstable DNA and lentiviral/retroviral vectors. Used by Addgene for pooled...
  13. Virus Protocol - Generating Stable Cell Lines

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    Protocol
    ...cells that have taken up the lentiviral transgene. Note that not all lentiviral vectors deliver antibiotic...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 ...associated with repeated transient transfection. Some lentiviral vectors deliver mammalian antibiotic resistance...
  14. Fluorescence Titering Assay

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    Protocol
    ...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...marker. Determining the titer of your lentiviral vector allows you to control the multiplicity of infection... 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 ...
  15. dTAG - You're it!

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    Blog Post
    ...available from Addgene. These plasmids enable lentiviral expression of N- or C-terminally FKBP12F36V-tagged...unclear which terminus should be tagged, the dTAG lentiviral plasmids can serve as a rapid means of comparing...multiple FKBP12F36V-tagged proteins using our lentiviral expression platform, to assess selective degradation...
  16. Hot Plasmids - October 2020

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    ... The Human GlycoGene CRISPR Library is a new lentiviral CRISPR library that targets human glycosyltransferase...inhibition pooled library is now available as lentiviral preps in addition to the pooled library format...library inhibits over 18,000 human genes. Find the lentiviral preps at Addgene hSyn1-SIO-stGtACR1-FusionRed...
  17. CRISPR 101: Epigenetics and Editing the Epigenome

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    ...Core, as well as pLV-dCas9-p300-P2A-PuroR for lentiviral expression. Figure 1: dCas9-p300 adds ...decrease methylation and activate transcription. A lentiviral vector with the same modifier, Fuw-dCas9-Tet1CD...Tet-dependent (TetO-dCas9-D3A) constructs. For lentiviral expression, Fuw-dCas9-Dnmt3a and Fuw-dCas9-Dnmt3a-P2A-tagBFP...hUbC-dCas9-MQ1(Q147L)-EGFP is also available for lentiviral transduction. Lysine-specific Demethylase 1 (...Jaenisch’s lab in plasmid form or as ready-to-use lentivirus. Figure 2: dCas9-Tet1 demethylates cytosines...with the former also available as ready-to-use lentivirus. Figure 3: dCas9-DNMT3A methylates cytosines...
Showing: 41 - 60 of 188 results