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We narrowed to 72 results for: AAV Cas9

Showing: 41 - 60 of 72 results

  1. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    Published
    Aug. 18, 2016, 2:30 p.m.
    ... 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...

  2. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    Published
    Feb. 17, 2022, 2 p.m.
    ...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...

  3. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    Published
    Jan. 26, 2016, 3:30 p.m.
    ...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...

  4. Hot Plasmids - February 2022

    Type
    Blog Post
    Published
    Feb. 22, 2022, 2:15 p.m.
    ...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...

  5. 27 Hot Plasmids from 2016

    Type
    Blog Post
    Published
    Dec. 22, 2016, 3:03 p.m.
    ...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...

  6. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    Published
    Aug. 29, 2024, 1:15 p.m.
    ...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...

  7. New CRISPR Web Resources and #12DaysofCRISPR Recap

    Type
    Blog Post
    Published
    Dec. 12, 2017, 3:13 p.m.
    ...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...

  8. Generating Mouse Models Using CRISPR/Cas9

    Type
    Blog Post
    Published
    July 12, 2016, 2:30 p.m.
    ...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...

  9. Hot Plasmids and Viral Preps - September 2021

    Type
    Blog Post
    Published
    Sept. 21, 2021, 1:15 p.m.
    ...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  ...

  10. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    Published
    Sept. 10, 2020, 11:45 a.m.
    ... 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...

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

    Type
    Blog Post
    Published
    April 24, 2015, 2:39 p.m.
    ...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...

  12. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    Published
    July 24, 2018, 12:51 p.m.
    ...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...

  13. How Addgene Creates Content

    Type
    Blog Post
    Published
    Nov. 17, 2022, 2:15 p.m.
    ...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...

  14. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    Published
    Jan. 6, 2015, 5:21 p.m.
    ...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...

  15. Prime Editing: Adding Precision and Flexibility to CRISPR Editing

    Type
    Blog Post
    Published
    Jan. 13, 2025, 9:49 p.m.
    ...prime editor consists of a Cas9 nickase fused to a reverse transcriptase. The Cas9 nickase is guided to the... that most Cas9-induced DSBs are repaired by non-homologous end joining. As a result, Cas9 treatment causes... ~ ~ ✓ ✓ ✓ R221K N394K Cas9 mutations Improve Cas9 nuclease activity ~ ~ ~ ~ ...homology to the genomic DNA locus. After nicking by Cas9, the homologous pegRNA sequence hybridizes to the...-MLV) reverse transcriptase to the C-terminus of Cas9 H840A nickase (PE1). But innovations by the Liu ...generate precise changes. However, the efficiency of Cas9 cleavage is relatively high while the efficiency...ratios of desired edits to indel byproducts than Cas9-initiated HDR. Prime editing tools Prime editing...

  16. Teaching an Old DOG New Tricks: Controlling Protein Activity with GFP

    Type
    Blog Post
    Published
    Nov. 24, 2015, 3:30 p.m.
    ...and 100% of DsRed+ cells also expressed GFP. Using AAV constructs, Tang et al. found that Cre-DOG could ...optogenetic studies, and since cells infected with AAV-Cre-DOG retain normal neuronal function, this system...versions of non-modular proteins like Cre. Could Cas9-DOG be developed soon to allow genome modification...

  17. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    Published
    May 14, 2024, 1:15 p.m.
    ...unique sgRNAs into a multi-TU targeting construct (Cas9, sgRNA, reporter, etc.). This allows for the multiplexing...repurposed for CRISPR applications (gene insert = Cas9 + gRNA), Addgene distributes specialized CRISPR ...and to compare different Cas enzymes in all-in-one AAV delivery. While Fragmid developers were specifically...Fragmid toolkit to facilitate other lentivirus- and AAV-based studies. Unified MoClo Systems While many MoClo...High-Efficiency Genome Editing in Plants Mediated by a Cas9 Gene Containing Multiple Introns.” Plant Communications...

  18. Viral Vectors 101: Parts of the AAV Transfer Plasmid

    Type
    Blog Post
    Published
    Aug. 11, 2020, 1:15 p.m.
    ...makes an AAV transfer plasmid an AAV transfer plasmid. ITR sequences are 145 bases each and AAV plasmids... want to do, but it requires some AAV. You’ve never worked with AAV before, but you aren’t going to let...experiments, making AAV starts with some plasmids. You just need three plasmids to start making AAV:  the packaging... the AAV transfer plasmid and take a look at each of its parts. Figure 1: Parts of an AAV transfer...   Key elements of an AAV plasmid First let’s talk about the key parts of an AAV plasmid. Inverted terminal...packaged into the AAV molecule. What’s outside of the ITRs does not get packaged in the AAV vector. The cloning...ITR loss abolishing AAV packaging. To prevent ITR loss, it’s best to propagate AAV constructs in recombination-deficient...

  19. Hot Plasmids - November 2023

    Type
    Blog Post
    Published
    Nov. 14, 2023, 3:35 a.m.
    ... ideal for size-limited AAV delivery. PE6c and PE6d are optimized for dual-AAV systems. These variants...edits, such as long or highly structured templates. Cas9 variants PE6e–g often had even higher editing efficiencies...

  20. Hot Plasmids - August 2020

    Type
    Blog Post
    Published
    Aug. 18, 2020, 1:15 p.m.
    ...syn-FLEX-axon-jYCaMP1s. Find these AAVs at Addgene Cre-dependent EYFP AAV in the new serotype PHP.V1 that...pEF1a-DIO-FLPo-WPRE-hGHpA, which is now also available in AAV5 and AAVrg. Find these AAVs at Addgene New controls, biosensor...plasmids at Addgene! CRISPR RNP electroporation and AAV donor infection (CRISPR-READI) enables genome engineering...transduction of vesicular brain cells. Find this AAV at Addgene New serotypes for the Cre-dependent ...biosensor, chemogenetic and optogenetic AAV for expression in forebrain GABA-ergic interneurons. These include...hDlx-GiDREADD-dTomato and mDlx-ChR2-mCherry. Find these AAVs at Addgene ...genome using AAV1 and contain an EGFP reporter. Expression of the dCas9-KRAB activator or dCas9-VPR repressor...
Showing: 41 - 60 of 72 results