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We narrowed to 314 results for: cas9 genes

Showing: 121 - 140 of 314 results

  1. dTAG - You're it!

    Type
    Blog Post
    ...and endogenous ENL was inactivated using CRISPR/Cas9 gene-editing. Featured molecules: dTAG-7 and dTAG-13... endogenous MELK was inactivated using CRISPR/Cas9 gene-editing. Featured molecules: dTAG-7, dTAG-13, ...CRISPR/Cas9-mediated knock-in. Featured molecule: dTAG-47 Additional Resources on the Addgene Blog Easi-CRISPR... Easi-CRISPR: Generating Knock-In and Conditional Mouse Models CRISPR 101: Cas9 Nickase Design and Homology...Plasmids for endogenous gene tagging in human cells Resources on Addgene.org Browse all CRISPR plasmids...selective degradation of FKBP12F36V-BRD4 using CRISPR/Cas9-mediated knock-in, and to evaluate in vivo degradation...Figure 1). Plasmids: We generated a series of plasmids that are available from Addgene. These plasmids enable...

  2. CRISPR 101: Homology Directed Repair

    Type
    Blog Post
    ...Cas-mediated Transgenesis (2018). Pubmed. CRISPR References: Genome engineering using the CRISPR-Cas9 system...2014). PubMed. The Use of CRISPR/Cas9, ZFNs, and TALENs in Generating Site-Specific Genome Alterations...Homology directed repair and genome engineering CRISPR-Cas9 has revolutionized the genome engineering world ...the gRNA/PAM site remains intact. In other words, Cas9 will keep cutting and repairing your target DNA ...of Genome Engineering Approaches with the CRISPR/Cas9 System. Li, K et al. PLoS ONE 9(8); (2014). PubMed...editing by catalytically active and inactive CRISPR-Cas9 using asymmetric donor DNA. Richardson CD, et al...your desired edit are all you need to generate a knock-in.   General considerations for designing a repair...

  3. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    ...CRISPR/Cas9 constructs. zCas9i is a high efficiency intron-optimized SpCas9-coding gene. SlugCas9 is a ... 0.001% of the population.  Because indel generation by Cas9 can occur at a steady rate, scientists found...promoters driving expression of Cas9 were used in this study.  The zCas9i cloning kit is a plant cloning...activity to SaCas9. A new yeast inducible CRISPRi library - Ampl133 targets S. cerevisiae genes with 6-12...a compact Cas9 ortholog from Staphylococcus lugdunensis that recognizes a NNGG PAM and has comparable ...12 guides per gene where gRNA expression is induced by anhydrous tetracycline. Three new anti-CRISPRs ...anti-CRISPRs were identified that inhibit the S. aureus Cas9, a Cas protein that is smaller and easier to deliver...

  4. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    ...isoform-specific gene expression. Gene isoforms refer to mRNAs generated from the same gene but with different...variant of Cas9 (dCas9) and engineered combinations of transcriptional activators to induce gene transcription...isoform-specific gene regulation — at least, more straightforward than generating cDNA for each gene isoform individually...first ask yourself whether gene overexpression is necessary (instead of gene knockout or knockdown, for...with neuron-specific expression of dCas9-VPR (Figure 2), a popular Cas9 activator developed by the Church... neuron-optimized CRISPR/dCas9 activation system for robust and specific gene regulation." eNeuro 6.1 ...transcriptional activation of multiple genes in the brain using CRISPR–dCas9-activator transgenic mice." Nature...

  5. What's New in CRISPR - Spring 2019

    Type
    Blog Post
    ... - this time using the Staphylococcus aureus Cas9 (SaCas9). This AAV-based genome editing system is regulated...the Cas12b plasmids  Plant base editing with a Cas9-adenosine deaminase fusion Caixia Gao’s lab has ...tRNA adenosine deaminase that is fused to a nicking Cas9. This base editor can be used for A•T to G•C edits... a few of the new CRISPR plasmids available at Addgene. We will still periodically focus on specific CRISPR... in Molecular Neuroscience Find the AAV-based SaCas9 genome editing plasmids String assembly gRNA ...is a single step gRNA multiplexing system that generates vectors containing high numbers of gRNAs in one...a new CRISPR tool you’ve recently deposited to Addgene and you’d like it to be included in the next What...

  6. Typing CRISPR Systems

    Type
    Blog Post
    ...domain due to a gene that resembles cas10. All type IV subtypes are missing adaptation genes, Cas1 and Cas2...–C, with the beloved SpCas9 (from Streptococcus pyogenes) belonging in II-A. Cas9 was the first to be ...sequence similarity, phylogenetic analysis, neighborhood analysis of surrounding genes, and experimental ...first time. The discovery and application of CRISPR-Cas9 kicked off over a decade of frenzied interest in...straightforward as it seems. There is not a single gene that is shared across all CRISPR types; the fast...while Class 2 use single protein Cas effectors, like Cas9. The Cascade complex doesn’t usually carry out the...together because they contain a distinct Cas7-type gene. The exact mechanism of type IV systems is not known...

  7. How to Design Your gRNA for CRISPR Genome Editing

    Type
    Blog Post
    ...hammer: Gene knockout by NHEJ Gene knockout with CRISPR technology is usually accomplished by Cas9-mediated...technologies – a nuclease-dead Cas9 (dCas9) is directed near the promoter of a target gene. Here, the target window...you are trying to do: gene knockout, a specific base edit, or modulation of gene expression. Location ...multiple sequences targeting a gene to score in order to call that gene a hit.  This is an exciting time...Agreement between two large pan-cancer CRISPR-Cas9 gene dependency data sets. Nat Commun 10: . https:/...protein-coding capacity of a locus. When using S. pyogenes Cas9 (SpCas9), potential target sites are both [5’-20nt-NGG...This means that, for gene editing, there are usually few potential gRNAs. While SpCas9, with a PAM preference...

  8. Design Tips for Prime Editing

    Type
    Blog Post
    ...editing technique that relies on a fusion enzyme of Cas9 nickase + reverse transcriptase and a prime editing...., 2019), disrupting its canonical structure and Cas9 binding (Nishimasu et al., 2014). Figure...pegRNAs that use other bases in this location bind to Cas9 with canonical sgRNA structure (right). Created ...along with your intended edit. This prevents the Cas9 nickase from re-binding and nicking the newly synthesized...Zhang, F., & Nureki, O. (2014). Crystal Structure of Cas9 in Complex with Guide RNA and Target DNA. Cell, ...and goal of the experiment. Now, onto the tips! General principles pegRNA design If you’re just starting...programs to help you design pegRNAs. Here on the Addgene blog, we featured a guest blog by the developer...

  9. CRISPR References and Information

    Type
    Collection
    ... sites for S. pyogenes Cas9, S. aureus Cas9, S. thermophilus Cas9, N. meningitidis Cas9, or Cas12a from...vectors using ssDNA oligos p201G Cas9 ; p201B Cas9 ; p201H Cas9 ; p201N Cas9 ; pUC gRNA Shuttle PDF, 500 KB...CRISPR RNA array: Cas9 (pX260) or Cas9 D10A (pX334) ; tracrRNA: Cas9 (pX330) or Cas9 D10A (pX335) PDF,... window) CRISPR/Cas9 reagent generation; gRNA design; HDR design; Deep sequencing Cas9: pMLM3613 dsDNA...CRISPR/Cas9 Knockout (MAGeCK). This tool can identify positively or negatively selected sgRNAs, genes, or...118 KB Church gRNA design and cloning for Cas9 orthologs Cas9 plasmids PDF, 107 KB Chen and Wente Zebrafish...vitro transcription, injection gRNA core ; Cas9 ; optimized Cas9 PDF, 66.8 KB Fujii gRNA design and cloning...

  10. Trends in CRISPR and SynBio Technologies

    Type
    Blog Post
    ...the dramatic spread of the Cas9 technology: "The rapid adoption of the Cas9 technology was also greatly...program are here. This was a great opportunity for Addgene to present our own data on plasmid deposits and...and distirbution for these fast moving fields.  Addgene is a global nonprofit plasmid repository. Over ...Over 2,000 labs have deposited plasmids to Addgene and we distribute over 130,000 plasmids in 2014. Thus,...combination of open-source distributors such as Addgene, as well as a number of online user forums such...Meeting Eric presented a poster with data comparing Addgene global deposit and distribution trends for Zinc...

  11. Zhang Lab CRISPR Page

    Type
    Collection
    ...Return to top Cas9 SAM transcriptional activation plasmids and screening library CRISPR/Cas9 Synergistic... of endogenous genes. It consists of three components: A nucleolytically inactive Cas9-VP64 fusion A sgRNA...of cells and nuclei. Return to top AAV - Cas9 mouse CRISPR-Cas9 is a versatile genome editing technology...constitutively expressing Cas9 knockin mice (Platt et al ., Cell 2014). In these mice the CRISPR-Cas9 system can be...constitutively Cas9-expressing mouse. Described here are AAV vectors that can be combined with Cas9 in a wide...are below. The Cas9 knockin mouse can be purchased from Jackson Labs: Cre-dependent Cas9 mouse Constitutively...cloning information: CRISPR-Cas9 mouse toolbox protocol 637.8 KB CRISPR-Cas9 Cre expression vectors for...

  12. Rett Syndrome: A History of Research and Therapeutic Outlooks

    Type
    Blog Post
    ... on therapeutic strategies that use gene replacement therapy, gene editing, MECP2 reactivation, RNA editing...Novartis and rebranded to Novartis Gene Therapies, and Taysha Gene Therapies, founded on Gray’s work, ...corrected by CRISPR/Cas9 in wild-type human iPSCs. Image modified from Le et al., 2019. Gene editing via ...the CRISPR/Cas9 system to both precisely insert the MECP2-R270X mutation into the MECP2 gene in wild-type...protein 2 (MECP2) gene, Rett Syndrome is a rare, progressive, neurodevelopmental genetic disorder that affects... CRISPR/Cas9-Mediated MECP2 Modifications in Human-Induced Pluripotent Stem Cells. Front Genet 10: . https...of Rett Syndrome research, the role of the MECP2 gene, and highlights some of the latest breakthrough ...

  13. CRISPR Guide

    Type
    Collection
    ... available with multiple Cas9 variants, including high fidelity Cas9s, Cas9s with various PAM requirements...dual nickases to generate a knockout or gene edit; or using Cas9 to generate large genomic deletions or ...strand SpCas9-HF1 - disrupt Cas9’s interactions with DNA phosphate backbone HypaCas9 - increase Cas9 proofreading...Plasmids: Purify Cas9 Alternatives for CRISPR Genome Engineering S. pyogenes Cas9 (SpCas9) is the most commonly...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 ...

  14. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    ...critical for Cas9 cleavage), thereby rendering the resulting construct resistant to Cas9 induced cleavage... introduced to endogeneous DNA through homology-directed repair (HDR) of a CRISPR/Cas9 induced double-...resistant to Cas9 cleavage. If a plasmid contains the template, site-directed mutagenesis can be used ...footnote; a similar approach can be used to identify Cas9/CRISPR-induced genome modifications. Occasionally... the mutagenesis reaction. Alternative approaches to site directed mutagenesis Point-mutagenesis is fairly...relevant in this age of CRISPR; site-directed mutagenesis generally applies to plasmids, but may also facilitate...modifications.   Site-directed mutagenesis of Plasmids. A) Generation of mutagenized plasmids. PCR primers...

  15. Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015

    Type
    Blog Post
    ...using bacterial phage vectors with CRISPR/cas9 sensitizing gene cassettes. Thse vectors are engineered to... tetracycline cas9 inducible knock-out system. His lab is also exploring the use of cas9 proteins from...were identified for 61 such diseases. Now single gene alleles have been identified for 4,163, over 50%...Lowe talked about his work on the 500+ human tRNA genes and, until now, we've overlooked regulatory fragments...may be involved in preventing silencing of self-genes. Dr. Mello presented a hypothesis that these pathways...involved in RNA based surveillance of repetitive gene families and provide a way to guide inheritance ... to replace the drug resistantance genes in the bacteria thus making the bacteria sensitive to known antibotics...

  16. CRISPR Challenges: Standardization and Homology Directed Repair

    Type
    Blog Post
    ...engineered Cas9 variant might work for your experiment. PITChing MMEJ as an Alternative Route for Gene Editing...Corn’s lab describe how a better understanding of Cas9 biophysics can help enhance HDR. CRISPR 101: Ribonucleoprotein...tool to help you design, pick, and close guides for Cas9 and Cpf1. This tool supports over 250 genomes, and...template to increase efficiency of insertion and gene replacement. Care to share your favorite CRISPR ...Blugene and I represented Addgene at the recent Keystone meeting on Precision Genome Editing with Programmable...hopes to evaluate genome editing pipelines and generate a common lexicon for genome editing studies. These...desired edit, and I want to make sure that I (and Addgene) don’t forget this in the future. Here’s a list...

  17. Addgene @ Keystone: Thoughts on Precision Genome Engineering and Synbio

    Type
    Blog Post
    ...the CRISPR-Cas9 system. Mehmet Fatih Bolukbasi from Scot Wolfe's lab described a chimeric Cas9 that is improving...CRISPR-Cas9 field – Jennifer Doudna, Feng Zhang, and Keith Joung. All Addgene depositors! Addgene was mentioned...versions of each, and Tim Lu even compared his "memory" gene codes to a Turing machine (also discussed in the...harnessed. Sharing CRISPR, synbio, & more via Addgene Addgene's addition to the poster session was a first...attendant called them, "long, skinny things") and generally exuding a very-tired-but-very-excited attitude...happens): Science is hard. Chris Voigt, another Addgene depositor, discussed some of the many challenges...That so many scientists are tackling this type of genetic engineering is a testament to how amazing the downstream...

  18. 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... Co-expressed Cas9 Cas9 System Selection PI pCRISPR 42875 Bacteria BsaI none S. pyogenes Chloramphenicol...meningitidis Joung Cas9 sgRNA vector 68463 Mammalian U6 none S. pyogenes Zhang Cas9/pTREX-n 68708 Other/...independent U6 promoters and Cas9 from CBh promoter. Ventura A CRISPR/Cas9 toolkit for multiplex genome...polycistronic glycine tRNA-gRNA genes (PTGs) and then PTG cassettes are assembled into a Cas9 expressing vector. Yang...none S. pyogenes Qi pDD162 (Peft-3::Cas9 + Empty sgRNA) 47549 C. elegans yes, cut S. pyogenes Goldstein...cut S. pyogenes mCherry Kuhn pU6-(BbsI)_CBh-Cas9-T2A-BFP 64323 Mammalian BbsI yes, cut S. pyogenes BFP Kuhn...

  19. CRISPR Plasmids - C. elegans

    Type
    Collection
    ...Co-expressed Cas9 Depositing lab Cas9 species = S. pyogenes (PAM = NGG) 47549 pDD162 (Peft-3::Cas9 + Empty ...need Cas9 plasmid Joung 46170 PU6::klp-12_sgRNA cU6 PCR (see paper) Transfection none, need Cas9 plasmid... none, need Cas9 plasmid Boxem 47941 pMB60 T7 BsaI In vitro transcription none, need Cas9 plasmid Boxem...NHEJ. ID Plasmid Gene/Insert Promoter PI Publication Activate Catalytically dead dCas9 fused to a transcriptional...transcription of a specific gene. Design your gRNA sequence to direct the dCas9-activator to promoter or ... plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert Promoter PI Publication...or regulatory regions of your gene of interest. If the plasmid that you choose does not also express a ...

  20. CRISPR Plasmids - Drosophila

    Type
    Collection
    ...Delivery Resistance Co-expressed Cas9 Depositing lab Cas9 species = S. pyogenes (PAM = NGG) 49410 pCFD3-dU6...repair (HDR). ID Plasmid Gene/Insert Promoter PI Publication Prime Edit Cas9 H840A nickase fused to a ...transcription Virmilion none, need Cas9 plasmid Bullock and Port 49330 pAc-sgRNA-Cas9 dU6 BspQI Transfection Puromycin..., need Cas9 plasmid Bullock and Port 51026 U6-BbsI-crRNA dU6 BbsI Transfection none, need Cas9 plasmid...template. ID Plasmid Gene/Insert Promoter PI Publication Activate Catalytically dead dCas9 fused to a transcriptional...transcription of a specific gene. Design your gRNA sequence to direct the dCas9-activator to promoter or ... plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert Promoter PI Publication...
Showing: 121 - 140 of 314 results