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We narrowed to 92 results for: AAV Cas9

Showing: 41 - 60 of 92 results

  1. Meet the New Addgene Board Members!

    Type
    Blog Post
    ...project to develop an AAV-based COVID-19 vaccine.  Vandenberghe’s work with AAVs led to many interactions...Therapeutics and is an Addgene depositor. KSQ uses a CRISPR-Cas9 platform to systematically test the function of ...was compelled to be part of that.” He’s deposited AAV constructs for gene delivery. In 2018, he interviewed...

  2. 10 Great Guest Posts We're Thankful For

    Type
    Blog Post
    ...action. AAV: A Versatile Viral Tool for Gene Expression in Mammals Adeno-associated viruses (AAVs) are commonly...Cepko’s Lab at Harvard Medical School describes how AAVs function and indicates important things to consider... editing - all influenced by an understanding of Cas9 biophysics! Casilio: An Adaptive, Multitasking “... system that uses modular RNA binding proteins, dCas9, and a modified gRNA to direct a variety of customizable...

  3. Plan Your Experiment

    Type
    Collection
    ...organism without a specific preferred mutation Cas9 (or Cas9 nickase) Single (or dual) gRNA targeting 5′ ...generating a point mutation or inserting a tag Cas9 (or Cas9 nickase); Base editor Single (or dual) gRNA ...concern, Cas9-gRNA ribonucleoprotein (RNP) complexes are advantageous due to the transient Cas9 expression... screens using CRISPR AAV transduction CRISPR elements are inserted into an AAV transfer vector and used...used to generate AAV particles (for details, see our AAV Guide ) ∼4.5 kb packaging limit (only compatible... RNA delivery of Cas9 and gRNA In vitro transcription reactions generate mature Cas9 mRNA and gRNA, which...generate transgenic embryos Cas9-gRNA ribonucleoprotein (RNP) complexes Purified Cas9 protein and in vitro transcribed...

  4. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    ... Has Been Used with AAV Resources on Addgene.org Find Cas9 Activators Find Cas9 Repressors Catch Up ...CRISPR/Cas9 is an enormously plastic tool and has taken the scientific world by storm. While Cas9 has been...required for Cas9-based activation. CRISPRa for genome-wide studies One of the best uses for Cas9 activators...context of the gene, Cas9 activators efficiently solve these problems.   Which Cas9 activator should I ...Best Cas9 Variant for Your Next Experiment Find out More about Genome-wide Screens with CRISPR/Cas9 Learn...there has also been significant work on constructing Cas9 transcriptional activators. These constructs allow...upregulation of essentially any gene by fusing mutants of Cas9 deficient in DNA cutting activity to a transcriptional...

  5. New CRISPR Tools: Cas7-11 and PASTE

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    Blog Post
    ...compatible with both AAV and adenovirus templates. The system, which is larger than Cas9, can also be used...systems have enzymes small enough to fit into a dual AAV system, albeit with lower knockdown efficiencies....PASTEsystem involves insertion of landing sites via Cas9-directed reverse transcriptases, followed bylanding...bylanding site recognition and integration of cargo via Cas9-directed integrases.   Use PASTE can be delivered...

  6. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    ...long period of time. High fidelity Cas9s such as eSpCas9 and SpCas9-HF should be explored to reduce off-target...culture, but Ran et al. have shown that CRISPR and AAV can be used together for postnatal genome editing...facets of CRISPR DMD therapy. Long et al. show that AAV-CRISPR does not cause obvious germline modification...especially stem cells 2. Remove any immunogenicity of the AAV vector Once delivery has been optimized, it’s important...Majoros WH, Reddy TE, Gersbach CA. Multiplex CRISPR/Cas9-based genome editing for correction of dystrophin... vivo genome editing using Staphylococcus aureus Cas9. Nature. 2015 Apr 9;520(7546);186-91. doi: 10.1038...Tabebordbar et al. used the shorter SaCas9 with AAV8 and AAV9, respectively. Although each study used slightly...

  7. Hot Plasmids - February 2022

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    Blog Post
    ...CRISPR-based genetic screening methods have primarily used Cas9 to knock out genes, researchers at the Genetic Pertubation... a single expression cassette is advantageous to Cas9’s larger expression requirements (Figure 1), but... guides. While two guides require two arrays for Cas9, two guides can be streamlined into a single array...-wide library capable of performing similarly to Cas9 screens in a compacted library. Overall, Cas12a’...CRISPR-Cas effectors and fusion proteins, such as Cas9 and Cas12a, creates challenges for in vivo delivery...529 amino acids, less than half the size of either Cas9 or Cas12a. Despite its small size, CasMINI is comparable...off-target effects, yet small enough to be packaged in AAV. A dCasMini-mediated adenine base editor allows for...

  8. CRISPR Plasmids - Empty gRNA Vectors

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    Collection
    ... plasmid contains Cas9 and if so, which function of Cas9. We have plasmids with Cas9 that can cut, activate...Expression System Cloning Enzyme Co-expressed Cas9 Cas9 System Selection PI pCRISPR 42875 Bacteria BsaI...BbsI)_CBh-Cas9-T2A-BFP 64323 Mammalian BbsI yes, cut S. pyogenes BFP Kuhn pU6-(BbsI)_CBh-Cas9-T2A-mCherry...meningitidis Joung Cas9 sgRNA vector 68463 Mammalian U6 none S. pyogenes Zhang Cas9/pTREX-n 68708 Other...pX601 61591 Mammalian/AAV BsaI yes, cut S. aureus Zhang pX602 61593 Mammalian/AAV BsaI yes, cut S. aureus...independent U6 promoters and Cas9 from CBh promoter. Ventura A CRISPR/Cas9 toolkit for multiplex genome...cassettes are assembled into a Cas9 expressing vector. Yang A Robust CRISPR/Cas9 System for Convenient, High-Efficiency...

  9. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...pDGE Dicot Genome Editing Kit contains Cas9, Cas9 nickase, and Cas9 activator expressing plasmids that can... 74307 pAAV-CMV-DIO-Magneto2.0-sNRPpA AAV 74306 pAAV-CMV-DIO-TRPV4-p2A-ferritin-sNRPpA AAV 74302 ...CRISPR-Cas9-based tool for targeting CpG methylation by fusing catalytically inactive Cas9 (dCas9) with...high-throughput CRISPR/Cas9 gene modification in Caenorhabditis elegans CRISPR/Cas9 technology has revolutionized...growing collection of dCas9-VPR plasmids, which already includes dSaCas9-VPR in an AAV transfer vector for...reported on a novel Cas9-based activator developed by the Church lab, termed “dCas9-VPR”, which facilitates...gRNA expressing cells were then transfected with a Cas9-expressing plasmid to generate a population of mutant...

  10. Making CRISPR Plasmids Using Fragmid

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    Blog Post
    ...depending on above selections.) CRISPR enzyme (Cas9 [sp], Cas9 [other], Cas12a, RNA-targeting) CRISPR mechanism...CMV-driven; CROPseq) Mammalian AAV pRDA_889 (ITRs from AAV2) Mammalian Transposon pRDA_...Mammalian or drosophila) Delivery type (lentivirus, AAV, transposon, Plasmid) Destination vector (dropdown...

  11. New CRISPR Web Resources and #12DaysofCRISPR Recap

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    Blog Post
    ...Delivery Methods and A Match Made in Heaven: CRISPR and AAV Day 6: Bacteria: CRISPR Methods for Bacterial Genome...and CRISPR Antimicrobials Day 7: Multiplexing and Cas9 variants: CRISPR 101: Multiplex Expression of gRNAs...Pooled libraries: Genome-Wide Screening Using CRISPR/Cas9 and Plasmids 101: NGS Quality Control for Pooled... CRISPR Targeting Specificity with eSpCas9, SpCas9-HF1, & HypaCas9 Day 10: Transcriptional regulation:...The PAM Requirement and Expanding CRISPR Beyond SpCas9 Day 8: Repair mechanisms: CRISPR 101: Non-homologous...

  12. Generating Mouse Models Using CRISPR/Cas9

    Type
    Blog Post
    ...alternative Cas9, from Streptococcus aureus, is as effective at editing mouse embryos as Cas9 from S. pyogenes...Gurumurty et al., 2016). AAV can also be used as a vector to deliver CRISPR/Cas9 either through injection...guest bloggers, Wenning Qin and Haoyi Wang. CRISPR/Cas9 is revolutionizing the mouse gene-targeting field...ever to manipulate the mouse genome using CRISPR/Cas9. Similar to the human genome, the mouse genome ...robust in performance. In its natural setting, CRISPR-Cas9 is an acquired immune system in bacteria and archaea...) of about 125 nt that specifies the target, the Cas9 endonuclease that creates the DNA double-strand ...model basics To create a mouse model, the gRNA, Cas9, and donor oligonucleotide or plasmid components...

  13. Cre-lox system

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    Collection
    ...107312 AAV-hSyn-mCherry-P2A-Cre-WPRE mCherry and Cre; expressed in neurons hSyn AAV Yang 107313 AAV-aCamkII-mCherry-P2A-Cre-WPRE-BGH-polyA...Cre-dependent Cas9: Cre-dependent Cas9 can be knocked in to a mouse at the Rosa26 locus to facilitate Cas9 mediated...Deisseroth 55635 pAAV-EF1a-sCre SCre EF-1 alpha AAV Deisseroth 55636 pAAV-EF1a-Cre Cre EF-1 alpha AAV Deisseroth...CamKII AAV Wilson 105553 pENN.AAV.hSyn.Cre.WPRE.hGH Cre hSyn AAV Wilson 105555 pENN.AAV.hSyn.Cre.hGH Cre ...51507 AAV pmSyn1-EBFP-Cre Cre-EBFP fusion; Expression in neurons. synapsin AAV Zeng 51904 paavCAG-iCre ...iCre CAG AAV Kim 55632 pAAV-Ef1a-mCherry-IRES-Cre Cre and mCherry coexpression EF-1 alpha AAV Deisseroth...EGFP-Cre fusion hSyn AAV Wilson 105545 pAAV.CMV.HI.eGFP-Cre.WPRE.SV40 EGFP-Cre fusion CMV AAV Wilson 105550 ...

  14. Hot Plasmids and Viral Preps - September 2021

    Type
    Blog Post
    ...available including: pAAV-hsyn-GRAB_DA2h in AAV9 New GRAB-DA and 5HT sensors New AAV tools to selectively...end-point sequencing assays. These plasmids use “Cas9-mediated adenosine transient reporter for editing...from the Dimidschstein Lab More GCaMP8 ready-to-use AAV preps  ...

  15. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ... encoding the gRNAs and Cas9. As with the Gersbach lab plasmids, multiple Cas9 variants are available:...subsequently inserted into a Cas9-containing destination vector based on pCAMBIA. Cas9 is plant-optimized, and...humanized Cas9, D10A nickase mutant (Cas9n), dCas9 (transcriptional repression), and Fok1-dCas9 (dimeric...options include humanized wt Cas9, dCas9 (transcriptional repressor), and dCas9-VP64 (transcriptional activator... then digested using BsmBI and ligated into Cas9 or dCas9-containing destination vectors. Destination ...the multiple gRNAs and Cas9 together into a single plasmid. Although only wt hCas9 entry vectors are supplied...target multiple sites! In this post, we'll cover Cas9 multiplexing options, but also check out our blog...

  16. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    ...adeno-associated viral vectors (AAV) available - AAV-EF1a-BbChT and AAV-EF1a-BbTagBY. These constructs ...associated with AAV; co-infection with both constructs produces a minimum of 8 colors. The use of AAV provides...CRISPR-Cas9 genome editing may be the hot new way to manipulate gene expression, but other gene manipulation...

  17. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    ...lentivirus, Adenovirus, and Adeno-associated virus (AAV). If you are wondering which virus fits you experiments...catalytically inactive (dead), human codon-optimized Cas9 under the control of Murine Stem Cell retrovirus...backbone from the Zhang lab expressing S. pyogenes Cas9 and a gRNA cassette. It can be used to produce a... (shRNA, miR-30-shRNA), mutation/editing (CRISPR-Cas9) or deletion (Cre), together with expression of ...and selection marker removal. CRISPR technology pdCas9-humanized (Plasmid #44246) Expression of a catalytically...

  18. How Addgene Creates Content

    Type
    Blog Post
    ...Addgene’s mission statement extends beyond plasmids, AAVs, and antibodies to our content, website, Data Hub...enable that process. Our repository of plasmids, AAVs, antibodies, and data allows scientists to easily...changes and require updating or retiring. CRISPR/Cas9, for example, has gone through rapid change in the...

  19. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...plasmids available! Interested in purifying Cas9? Check out pET-28b-Cas9-His from Alex Schier's lab, designed...the lab of Yonglong Chen, pCS2-3xFLAG-NLS-SpCas9-NLS is a Cas9 expression plasmid that was used by Chen...colleagues, FokI-dCas9 expresses Fok1 nuclease domain fused to catalytically inactive Cas9 DNA-binding domain...been deposited to Addgene, including lentiviral and AAV expression vectors for certain variants. Klapoetke...gene/region of interest is a key step for CRISPR/Cas9 gene editing. Dr. Masahito Ikawa has created a GFP...Next, the gRNA being tested is expressed along with Cas9. If the gRNA effectively cuts the target sequence...designed for expression and purification of Cas9 protein in Rosetta E. coli. New Lentiviral CRISPR activator...

  20. Plan Your Experiment

    Type
    Guide
    ...organism without a specific preferred mutation Cas9 (or Cas9 nickase) Single (or dual) gRNA targeting 5′ ...generating a point mutation or inserting a tag Cas9 (or Cas9 nickase); Base editor Single (or dual) gRNA ...concern, Cas9-gRNA ribonucleoprotein (RNP) complexes are advantageous due to the transient Cas9 expression... screens using CRISPR AAV transduction CRISPR elements are inserted into an AAV transfer vector and used...used to generate AAV particles (for details, see our AAV Guide ) ∼4.5 kb packaging limit (only compatible... RNA delivery of Cas9 and gRNA In vitro transcription reactions generate mature Cas9 mRNA and gRNA, which...generate transgenic embryos Cas9-gRNA ribonucleoprotein (RNP) complexes Purified Cas9 protein and in vitro transcribed...
Showing: 41 - 60 of 92 results