We narrowed to 92 results for: AAV Cas9

...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...

...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...

...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...

...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...

...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...

... 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...more Cas9 fusion tools later in the guide. Figure 3: Overview of Cas9n. Increasing fidelity of Cas9 Researchers...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...

...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...

...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...

... 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...

...). You will decide how to express Cas9, the delivery system for Cas9 and the guide RNA (gRNA), the design...non-homologous end joining (NHEJ) at the cut site Cas9 or Cas9 nickase High-fidelity Cas enzymes increase specificity...insertions, by copying from a DNA donor template Cas9 or Cas9 fusions with proteins that promote HDR Requires...site. Consider alternative Cas9 enzymes with different PAM sequences or Cas9 enzymes with flexible PAM ... collection of in-stock lentiviral preps . AAV vectors AAV vectors offer some advantages for viral vector...delivery. Both lentiviral and AAV vectors are safe to use in the lab, but AAVs exhibits the lowest immunogenicity... transgene expression with AAVs can be long-lived, despite recombinant AAVs not integrating into the host...

...about how to minimize leakiness in experiments using AAV vectors. Highlighted Plasmids Browse the tables below...preps feature Cre-dependent tools, or consider our AAV Packaged on Request service for more options. Addgene...Activity with GFP Plasmids 101: FLEx Vectors Optimizing AAV DIO and FLEx Vector Expression Plasmids 101: Knockout...preps feature Flp-dependent tools, or consider our AAV Packaged on Request service for more options. Addgene...CRISPR Knockout Library CRISPR Sidi Chen Use this AAV CRISPR knockout library expressing luciferase and...vivo using a mouse line expressing Cre-dependent Cas9. Kits Kit Name Type PI Description dRMCE Plasmid...Return to top Related Addgene Plasmid Collections AAV Enhancer Collection Fluorescent Biosensors Chemogenetics...

...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...

...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...

...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...

...APEX targeting Alice Ting 85040 pK170.AAV-TRE-Cre-WPRE (Supernova) AAV vector for dox-inducible expression...sgRNA TetR H1-2O2 Nathanael Gray 35625 pAAV-Ptet-RFP-shR-rtTA AAV Tet-On shRNA vector. To evaluate shRNA...promoter Tet-On 3G rtTA David Vereide 120309 pAAV-FAH-rtTA3G AAV vector to express Tet-On 3G transactivator...promoter TetR Eric Campeau , Paul Kaufman 175274 pAAV-rtTA AAV Tet-On vector with neuron-specific expression...Plasmid #27106 for rtTA. tTA Edward Hsiao 99118 pAAV-CAG-tTA AAV Tet-Off vector, expresses tTA from the CAG... Tet-Off tTA-Advanced Takeshi Imai 104109 pAAV-Syn1-tTA AAV Tet-Off vector, expressed tTA from the hSyn...tet-inducible Cas9-2A-EGFP expression. Based on LentiCRISPR v2 . Adam Karpf 167935 pLenti-tetON-KRAB-dCas9-DHFR-EF1a-TagRFP...

...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 ...

... 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...

...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...

...pX330S-2-PITCh - expresses Cas9 & PITCh-gRNA pX330A-FBL/PITCh - expresses Cas9, PITCh-gRNA, & FBL-specific...#79888) , which expresses an untagged Cas9 AND a gRNA targeting the AAVS1 locus. Doyon Tagging Plasmid: 3xFLAG... consists of two components: A vector containing Cas9 and a validated gRNA, based on the Zhang lab's PX458...need to design and clone in a gRNA(s) to guide the Cas9 protein to your target sequence, as well as design...consists of two components: An all-in-one CRISPR/Cas9 vector based on the Zhang lab's PX330 , simultaneously... to generate both an expression cassette for the Cas9-programming sgRNA and HR donors for selectable genome...for integrating at the AAVS1 "safe harbor" locus: eSpCas9(1.1)_No_FLAG_AAVS1_T2 Doyon Lab TAP Tagging ...

...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...