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We narrowed to 92 results for: AAV Cas9

Showing: 61 - 80 of 92 results

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

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ...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...

  3. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    ...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...

  4. CRISPR History and Development for Genome Engineering

    Type
    Collection
    ...High fidelity Cas9s: Three high fidelity Cas9 variants , SpCas9-HF, eSpCas9, and HypaCas9 display very ...Staphylococcus aureus (SaCas9) is smaller than SpCas9 and more easily packaged in AAV. Cas9 orthologs also have...III and Cas9 enzymes, which together separate the individual crRNAs. (3) The tracrRNA and Cas9 nuclease...beyond SpCas9, moving us closer to being able to target every locus in every genome. Multiple Cas9 orthologs...capability of Cas9 to make other types of modifications at specific loci. Activate or Repress : dCas9 fused to...the crRNA would bind. (5) After the complex binds, Cas9 separates the double stranded DNA target and cleaves...Single multi-domain effector) Type I (Cas3) Type II (Cas9) Type III (Cas10) Type V (Cpf1) Type IV (Csf1) Type...

  5. CRISPR Plasmids - Prime Edit

    Type
    Collection
    ...contains Cas9. ID Plasmid Expression System Promoter Guide RNA Type Cloning Enzyme Co-expressed Cas9 Selection...transcriptase (RT) is fused to the C terminus of Cas9 H840A nickase. The fusion enzyme is capable of installing...template. During prime editing, the pegRNA directs the Cas9 nickase to the target sequence where it nicks the...nuclear localization signals, and two mutations in Cas9 to enhance nuclease activity; can be used in PE2...PE6 - small prime editors with optimized RT and/or Cas9 domains PE7 - addition of RNA-binding exonuclease...PuroTK-pegRNA_GG Mammalian, piggyBac hU6 pegRNA BsmBI Yes (Cas9 H840A + MMLV RT) PuroTK Jacob Giehm Mikkelsen 174038... RNA-H1-nick sgRNA-mCherry Mammalian, AAV hU6 + H1 pegRNA + nicking sgRNA No mCherry Hyongbum...

  6. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...adeno-associated virus (AAV) integration sites in AAV virions during AAV wild-type and recombinant AAV vector production...both key advantages of rAAV vectors. AAV Types Serotypes AAV serotypes are AAV variants that differ in...Parts of the AAV Transfer Plasmid Parts of the AAV Packaging Plasmid Producing Your rAAV AAV Serotypes and...Adeno-Associated Viral (AAV) Vector Guide Jump To: AAV Vectors Vector Production Integration AAV Types Common Uses...Services Packaged-on-Request AAV Preps In-Stock AAV Preps Resources on addgene.org AAV Plasmids Available at ...Virus (AAV) for Cell and Gene Therapy Adeno-associated Viruses (AAVs) for Genome Editing Using AAV for Neuronal...More AAV Vector topics Addgene Protocols General Transfection AAV Production in HEK293T Cells AAV Purification...

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

    Type
    Blog Post
    ...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...

  8. Neurodegeneration Research Collection

    Type
    Collection
    ... Target neural oxytocin receptors using an AAV-CRISPR/Cas9 strategy for gene editing across divergent ... Nat Biotechnol. 2023 May 22. See More AAV Viral Preps Find AAV viral preps for systemic delivery of viral...research. Disease Info Plasmid Collection CRISPR Tools AAV Viral Preps iPSC Differentiation Factors Antibodies...species. Boender et al. Sci Adv. 2023 Jun 2. Use Cas9 in astrocytes. Endo et al Science. 2022 Nov 4. Base...brain cells and more. New and Noteworthy: AAV.rTH.PI.Cre.SV40 ready-to-use viral prep useful for dopamine...

  9. Hot Plasmids - November 2023

    Type
    Blog Post
    ... 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...

  10. Hot Plasmids - August 2020

    Type
    Blog Post
    ...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...

  11. Empty Backbones - Choosing Your Perfect Plasmid Backbone

    Type
    Collection
    ...protein production AAV-GfaABC1D-MCS--4x6T-WPRE - For astrocyte-selective expression in AAV Dual promoter Separate...Advantages Representative Empty Backbones AAV High transduction efficiency, but do NOT integrate...expression, see our dedicated Adeno-associated Virus (AAV) Plasmids page Lentiviral Can transduce both dividing... Separate expression pRGEB31 - Deliver sgRNA and Cas9 into plants by Agrobacterium-mediated transformation...Express sgRNA/PTG with rice snoRNA U3 promoter and Cas9 with rice ubiquitin promoter for Agrobacterium-mediated... FP Collection NLS Nuclear localization dCas9 plasmid - dCas9 expression plasmid without effector fusions...

  12. Adenovirus Guide

    Type
    Guide
    ...for replication. For more information about AAVs, read our AAV guide . Recombinant Adenoviral Vectors Wild-type...big to be packaged in rAAV vectors. This is the case of some commonly used Cas9 homologs, especially when...double-stranded DNA genome. Adeno-associated viruses (AAV), however, are small single-stranded DNA viruses ...viruses that belong to the Parvoviridae family. AAVs were discovered as a contaminant of adenovirus preparations...how they got their name, but they are not related. AAVs require the presence of adenoviral genes E1, E4,... or when needing to package larger cargo. CRISPR-Cas9 Delivery As compared to other viral vectors, rAdV...vectors. This capacity is large enough to carry the Cas9 gene and a gRNA expression cassette in one single...

  13. CRISPR Guide

    Type
    Collection
    ... available with multiple Cas9 variants, including high fidelity Cas9s, Cas9s with various PAM requirements...strand SpCas9-HF1 - disrupt Cas9’s interactions with DNA phosphate backbone HypaCas9 - increase Cas9 proofreading...new tools. Figure 3: Overview of Cas9n Nickase and dead Cas9 The two Cas9 nuclease domains, RuvC and HNH...enhanced specificity by engineering Cas9 into high fidelity Cas9s (hfCas9) . There is currently no defined...application. Cas9 Orthologs While SpCas9 is the most commonly used Cas enzyme, additional Cas9 orthologs ...constructs with Cas9 fused to proteins that modulate HDR or NHEJ components, like miCas9 , Cas9-DN1S , or others...Plasmids: Purify Cas9 Alternatives for CRISPR Genome Engineering S. pyogenes Cas9 (SpCas9) is the most commonly...

  14. Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2

    Type
    Blog Post
    ...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 ...

  15. 8 Great Depositor Research Stories

    Type
    Blog Post
    ... 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...

  16. CRISPR Guide

    Type
    Guide
    ... available with multiple Cas9 variants, including high fidelity Cas9s, Cas9s with various PAM requirements...strand SpCas9-HF1 - disrupt Cas9’s interactions with DNA phosphate backbone HypaCas9 - increase Cas9 proofreading...new tools. Figure 3: Overview of Cas9n Nickase and dead Cas9 The two Cas9 nuclease domains, RuvC and HNH...enhanced specificity by engineering Cas9 into high fidelity Cas9s (hfCas9) . There is currently no defined...application. Cas9 Orthologs While SpCas9 is the most commonly used Cas enzyme, additional Cas9 orthologs ...constructs with Cas9 fused to proteins that modulate HDR or NHEJ components, like miCas9 , Cas9-DN1S , or others...Plasmids: Purify Cas9 Alternatives for CRISPR Genome Engineering S. pyogenes Cas9 (SpCas9) is the most commonly...

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

    Type
    Blog Post
    ...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...

  18. Distribution to Industry

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    Collection
    ...Carrier Proteins (SLCs) from the RESOLUTE collection AAV Packaging from Addgene Parkinson's Research from ...Stiewe Use these donor vector libraries for CRISPR/Cas9-mediated saturation genome editing of human TP53...

  19. What's New in CRISPR - March 2020

    Type
    Blog Post
    ... 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...

  20. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    ...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...
Showing: 61 - 80 of 92 results