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We narrowed to 72 results for: AAV Cas9

Showing: 61 - 72 of 72 results

  1. Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2

    Type
    Blog Post
    Published
    May 26, 2020, 1:15 p.m.
    ...contains Cas9. The Madison lab also created an inducible SRIRACCHA system (iSRIRACCHA) where Cas9 expression... labor intensive and quite time consuming.   As Cas9/gRNA activity is crucial for successful CRISPR editing...the scientific community for efficient real-time Cas9 activity assays. Recently scientists have created...fluorescent reporter assays to quantify targeted Cas9 cleavage, and repair through homology directed repair...containing the reporter gene that was cleaved by Cas9 and repaired via HDR using the supplied donor plasmid...expressing cells provides a real-time read out of Cas9 activity.  By using the RFP:GFP ratio, edited cells...Thus SRIRACCHA enables scientists to both quantify Cas9 activity and sgRNA efficiency as well as enrich ...

  2. 8 Great Depositor Research Stories

    Type
    Blog Post
    Published
    Dec. 15, 2022, 2:15 p.m.
    ... to use dCas9 (with no modifications!) as a CRISPR/dCas9-based tool for DNA demethylation. AAV Q&A with... excellent Q&A from Tim Miles on everything from AAV prep to targeting to animal models. It was one of...ancestral bias in CRISPR guide library design.  dCas9-directed DNA Demethylation: Guest blogger Daniel...

  3. CRISPR 101: Cas9 Nickase Design and Homology Directed Repair

    Type
    Blog Post
    Published
    March 15, 2018, 12:59 p.m.
    ...strands bluntly with WT Cas9 and one gRNA, you can create a staggered cut using a Cas9 nickase and two gRNAs...CRISPR nickase experiment Overview of Cas9 nickase We’ll use SpCas9 nickases as examples for this post. ...By mutating one of two Cas9 nuclease domains, researchers created the CRISPR nickase. Nickases create...targeting range: using an individual gRNA with WT Cas9, repair levels decrease rapidly 10 bp from the cut...The HDR efficiency compared favorably to using WT Cas9 with the left gRNA across the window. In a subsequent... this problem. D10A nickase outperformed both WT Cas9 sites tested by a wide margin, exhibiting >20% repair...adapted from an IDT webinar: Optimized methods to use Cas9 nickases in genome editing. For more information...

  4. What's New in CRISPR - March 2020

    Type
    Blog Post
    Published
    March 10, 2020, 1:15 p.m.
    ... lower average Cas9-independent off-target DNA editing, ~5–50-fold lower levels of Cas9-dependent off-...Caulobacter genes using a catalytically dead Cas9 (dCas9) derived from Streptococcus pasteurianus  or ...s lab identified and validated CBEs with reduced Cas9-independent deamination. They generated these editors...generated hPSC lines with AAVS1-integrated, inducible, and fluorescent dCas9-KRAB and dCas9-VPR for quantifying...Saccharomyces cerevisiae. The system consists of dCas9, the mammalian transcriptional repressor Mxi1, and...accessible and the complex enters the nucleus where dCas9-Mxi1 can repress gene expression. In the dark, the...Caulobacter cells for ctrA gene expression using various dCas9. Image adapted from Guzzo et al., 2020. Caulobacter...

  5. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    Published
    Feb. 16, 2023, 7:05 p.m.
    ...out the gene for your protein of interest. CRISPR-Cas9 and a custom gRNA is all you need to generate a ...interference (CRISPRi) targets a catalytically dead Cas9 fused to a transcriptional repressor to your gene...) are transcriptional activators fused to a dead Cas9. The system is led, via a custom gRNA, to your gene...protein levels. Looking for a DD? Check out our DD-Cas9 vector! The price of tunable control These systems...Plasmid 101: Repressible Promoters CRISPR 101: Which Cas9 Do I Choose for my Experiment Additional resources...? Check out Addgene's Lentiviral dCas9 SAM vector and Piggybac dCas9 SAM vector! On/off control all the...repression of protein expression. Addgene has a dCas9 KRAB plasmid for transient knockouts!  Turn it on...

  6. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    Published
    Dec. 23, 2015, 3:30 p.m.
    ...fully functional Cas9 upon dimerization. Splitting wild-type Cas9 into n-terminal (Cas9(N)-2xNLS) and c-terminal... can write a blog post about it! Split Cas9 system The Cas9 protein is composed of an n-terminal DNA... the bi-lobed architecture of Cas9 to engineer a series of "split" cas9 molecules that cannot function...activation, the c-terminal cas9 fragment was fused with FK 506 binding protein 12 (Cas9(C)-FKBP-2xNLS) and the... the n-terminal cas9 fragment with FKBP rapamycin binding domain of mTor (Cas9(N)-FRB-NES) resulting in...rapamycin-inducible Cas9 for genome editing. Without rapamycin treatment, the Cas9(N)-FRB-NES fragment...sequence. Treatment with rapamycin induces Cas9(N)-FRB-NES and Cas9(C)-FKBP-2xNLS dimerization and net influx...

  7. Typing CRISPR Systems

    Type
    Blog Post
    Published
    March 18, 2025, 1:15 p.m.
    ...–C, with the beloved SpCas9 (from Streptococcus pyogenes) belonging in II-A. Cas9 was the first to be ...first time. The discovery and application of CRISPR-Cas9 kicked off over a decade of frenzied interest in...while Class 2 use single protein Cas effectors, like Cas9. The Cascade complex doesn’t usually carry out the...type VI, each relying on a unique Cas effector — Cas9, Cas12, or Cas13. If you’d like to explore Class...most popular and well-known type, as this is where Cas9 falls. The mechanism of type II is, unsurprisingly... Figure 3: Schematic of the basic CRISPR-Cas9 mechanism, representing type II-A. Created with ...effectors and is one of the most popular alternatives to Cas9. Cas12 is unique because it can process multiple...

  8. Hot Plasmids: Summer 2024

    Type
    Blog Post
    Published
    Aug. 27, 2024, 1:15 p.m.
    ...are much smaller proteins than SpyCas9, allowing them to be packaged in AAV vectors, potentially with multiplexed...from Neumann et al. 2024. The team showed that AAV-delivered CHARM could silence the expression of prion... (2024). Brainwide silencing of prion protein by AAV-mediated delivery of an engineered compact epigenetic... a programmable epigenetic editor consisting of dCas9 fused to a DNA methyltransferase — that can drive...DNA-targeting proteins, so the team created versions using dCas9, zinc finger proteins, and transcription activator-like...histone tail (H3K4me0) to a DNA-binding protein (dCas9 shown here). D3L recruits the endogenous, autoinhibited...

  9. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    Published
    Feb. 23, 2016, 3:30 p.m.
    ...containing Cas9 and the PITCh-gRNA. Step 3: Contransfect the PITCh vector with the vector carrying Cas9 and ...plasmic is cotransfected with a plasmid carrying Cas9, the PITCh-gRNA, and the locus-specifc gRNA. The...MMEJ-assisted gene knock-in using TALENs and CRISPR-Cas9 with the PITCh systems.” Nat Protoc. 11(1): 118-...advantageous to adapt PITCh to insert genes into AAVS1, the “safe harbor locus” of the human genome, as...

  10. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    Published
    March 21, 2017, 2:30 p.m.
    ...vectors fantastic options for delivering shRNA, CRISPR/Cas9 components, and fluorescent sensors. Retroviral...Viral Transductions Learn Why You Might Want to Use AAV in Your Research Lentiviral Vector Uses and Overview...

  11. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    Published
    Feb. 27, 2024, 2:15 p.m.
    ...cloned into it. gRNAs are used for directing CRISPR/Cas9 to a targeted genomic location, while shRNAs are...in bacteria) such as lentiviral, retroviral, and AAV plasmids, we recommend the NEB Stable strain. For...

  12. Hot Plasmids and Viral Preps - July 2021

    Type
    Blog Post
    Published
    July 27, 2021, 1:15 p.m.
    ...of GCaMP8 AAV preps has expanded. Check out the new selection of ready-to-use AAV9 and AAVrg vectors. ...available in AAV8! Expand your imaging experiments with the Cre-OFF/FLP-ON vector 137128-AAV8: Ef1a-Coff...editing in moderate thermophiles.  These evolved SpCas9 variants have improved activity on NAG PAMS and...
Showing: 61 - 72 of 72 results