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We narrowed to 314 results for: Cas9

Showing: 141 - 160 of 314 results

  1. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    ...containing Cas9 and the PITCh-gRNA. Step 3: Contransfect the PITCh vector with the vector carrying Cas9 and ...plasmic is cotransfected with a plasmid carrying Cas9, the PITCh-gRNA, and the locus-specifc gRNA. The...MMEJ-assisted gene knock-in using TALENs and CRISPR-Cas9 with the PITCh systems.” Nat Protoc. 11(1): 118-...

  2. CRISPR Plasmids - Drosophila

    Type
    Collection
    ...Enzyme(s) Delivery Resistance Co-expressed Cas9 Depositing lab Cas9 species = S. pyogenes (PAM = NGG) 49410...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...Plasmid Gene/Insert Promoter PI Publication Prime Edit Cas9 H840A nickase fused to a reverse transcriptase (...Injection or in vitro transcription Virmilion none, need Cas9 plasmid Bullock and Port 49411 pCFD4-U6:1_U6:3tandemgRNAs...pU6-BbsI-chiRNA dU6 BbsI Transfection none, need Cas9 plasmid O'Connor-Giles , Harrison , Wildonger 49408...Injection or in vitro transcription Virmilion none, need Cas9 plasmid Bullock and Port 49409 pCFD2-dU6:2gRNA dU6...

  3. Hot Plasmids September 2018 - Optogenetics, RNA Localization, Fluorescent Protein, and Base-editing Tools

    Type
    Blog Post
    ...reporter for rapid quantification of base-editing and Cas9 cleavage activity Article contributed by Angela ...lentiviral vector system that monitors both APOBEC- and Cas9-mediated Editing (ACE) in real-time. The reporter...they show that the ACE reporter can also quantify Cas9 cleavage, thus exhibiting its versatile utility ...Harris lab showed that human APOBEC3A-Cas9n-UGI and APOBEC3B-Cas9n-UGI base editing complexes are more ...fluorescence-based quantification of base editors that utilize dCas9 fused to cytidine deaminases like APOBEC. To achieve...and mCherry fluorescence is activated when APOBEC-Cas9n-UGI-mediated activity edits out a frame-shift mutation...

  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 ...III and Cas9 enzymes, which together separate the individual crRNAs. (3) The tracrRNA and Cas9 nuclease...Staphylococcus aureus (SaCas9) is smaller than SpCas9 and more easily packaged in AAV. Cas9 orthologs also have...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. Lentiviral Prep Service

    Type
    Collection
    ...Root Cas9 Lentivirus Cas9 viruses can be used to interrogate the genome in a variety of ways. Cas9 proteins... 52962 lentiCas9-Blast Sp Cas9 Cut Blasticidin Expresses human codon-optimized S. pyogenes Cas9 protein...backbone. Zhang Cas9 and Accessories for Activating Gene Expression Catalytically-dead Cas9 (dCas9) can be fused...plasmids resource page . Cas9 for Editing Genomic Sequences To perform genome edits, Cas9 nuclease is used to...Browse In-Stock Lentivirus Pooled CRISPR Libraries Cas9 Pooled Barcoding Libraries Control Addgene's lentiviruses...Resistance Activity PI 61422 dCAS9-VP64_GFP dCAS9 (D10A, H840A) none Expresses dCAS9-VP64 activator with 2A ...-targeting controls. This backbone contains SpCas9. SpCas9 and 76,441 unique sgRNAs targeting 19,114 human...

  6. Worm Expression Resources

    Type
    Collection
    ... via a CRISPR-Cas9 system. Cloning-free CRISPR for C. elegans , which uses purified Cas9, such as that...feeding worms bacteria expressing dsRNA or use CRISPR/Cas9 gene editing to easily engineer specific transgenic...worm CRISPR resources include plasmids expressing Cas9, empty gRNA backbones, and constructs for endogenous...targeting vectors for tag insertion using CRISPR/Cas9 and the selection of C. elegans strains. CRISPR ...that expressed from pHO4d-Cas9 from Michael Nonet. Protocol can be found at the (Link opens in a new window...Recovery of Custom Genetic Modifications with CRISPR/Cas9 in Caenorhabditis elegans. Mos1 Based Transgenesis...Blog Posts An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency Fluorescent Tagging of Endogenous...

  7. Cancer, Inflammation and Immunity - Harnessing the Body’s Defenses to Fight Cancer

    Type
    Blog Post
    ...in immunotherapy is the combination of the CRISPR/Cas9 methodology with T cell receptor engineering (Ref...using randomly integrating viral vectors, CRISPR/Cas9 genome editing allows targeted engineering of the.... "Targeting a CAR to the TRAC locus with CRISPR/Cas9 enhances tumour rejection." Nature 543.7643 (2017...

  8. Alex Chavez on MD PhDs, CRISPR, Gene Drives, & More

    Type
    Blog Post
    ...Advisory Board with a particular interest in generating Cas9 based tools to enable facile control of DNA and ... Addgene Blog Learn More About Alex's Work with Cas9 Activators Michael Koeris Explains His Path from...

  9. Plan Your Experiment

    Type
    Collection
    ...). 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 .../or Cas9 expression or (2) a lack of efficient target cleavage in cells that do express both Cas9 and ...edits without double-strand breaks Base editor (dCas9 or Cas9 nickase fusions with different base editing ...We will focus on making single edits using CRISPR/Cas9 in mammalian cells as an example, but many of these...

  10. Plant Plasmids and Resources

    Type
    Collection
    ... an intronized Cas9 series of vectors and DNA modules for assembly of CRISPR/Cas9 constructs - Sylvestre...CRISPR/Cas9 assembly for fast generation of multiple Arabidopsis mutants - Niko Geldner. CRISPR-Cas9 vector...plasmids expressing different functional variants of Cas9 and empty gRNA backbones. A few examples of useful...truncatula , wheat, and barley - Daniel Voytas. A CRISPR/Cas9 Toolkit for use in a variety of plant species (e.g...or more gRNA expression cassettes - Qi-Jun Chen. Cas9 and gRNA backbone for use in rice. Includes a protocol...species and genotypes - Zhenghe Li. A robust CRISPR/Cas9 system for convenient, high-efficiency multiplex...MoClo-compatible zCas9i Cloning Kit includes a high efficiency intron-optimized SpCas9-coding gene, zCas9i, and an...

  11. CRISPR Plasmids - Base Edit

    Type
    Collection
    ...are created by fusing Cas9 nickase or catalytically inactive “dead” Cas9 (dCas9) to a cytidine deaminase...editors are available with multiple Cas9 variants including high fidelity Cas9’s. Further advancements have ...base editors, the evolved TadA domain is fused to a Cas9 protein to create the adenine base editor. Both ... lab and CRISPR-X from the Bassik lab , in which Cas9 is fused to activation-induced cytidine deaminase...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...

  12. CRISPR-based Genome Editing Systems For Editing in E. coli

    Type
    Blog Post
    ...consists of one plasmid that expresses the S. pyogenes Cas9, a temperature-sensitive replicon for self-curing...), which we presume caused pTargetF to be cut by Cas9. This would result in the lack of recovered transformants...Editing in the Escherichia coli Genome via the CRISPR-Cas9 System. Appl Environ Microbiol 81:2506–2514 . https...2021) A modified pCas/pTargetF system for CRISPR-Cas9-assisted genome editing in Escherichia coli. Acta...edits. Once you have all the edits you want, cure pCas9 by growing the cells at 37 °C.     The new pEcCas...  The updates involved the following changes. In pCas9, we 1) replaced the promoter of gRNA-pMB1 with PrhaB...

  13. CRISPR Plasmids - Xenopus

    Type
    Collection
    ...Enzyme(s) Delivery Resistance Co-expressed Cas9 Depositing lab Cas9 species = S. pyogenes (PAM = NGG) 51306...designed for use in Xenopus. Cut Fully functional Cas9 enzymes designed to introduce a double-strand break...surrounding the DSB is introduced along with the Cas9 and gRNA plasmids, the cell may instead repair the...backbone T7 BsaI In vitro transcription none, need Cas9 plasmid Chen Do you have suggestions for other plasmids...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...

  14. Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering

    Type
    Blog Post
    ...than other Cas13s and a whopping 33% smaller than Cas9! Their small size makes then easy to package in ...significant off-targets, CasRx displayed zero. Unlike Cas9, for which efficiency varies widely across guide...CRISPR RNA Editing with REPAIR Get tips for using Cas9 Transcriptional Activators Learn about Cas13a (C2c2...regulating methods: small hairpin RNA interference, dCas9-mediated transcriptional inhibition (CRISPRi), and...

  15. CasPEDIA: A Functional Classification of Cas Enzymes

    Type
    Blog Post
    ... CRISPR 101: Which Cas9 do I choose for my CRISPR experiment? CRISPR 101: Cas9 The Other Cas(s) The...Addgene. So whether you're looking for the exact Cas9 plasmid used by Jinek et al. 2012, or browsing for...Once upon a time, not so long ago, spCas9 was the only Cas enzyme widely available and applied by researchers...and trans activity of each entry. For example, SpyCas9a (one of the most frequently used Cas enzymes) ...CasPEDIA Cas ID display information for a given Cas (SpyCas9a) from CasPEDIA.   Helpful resources and information...List of resources and properties for a Cas entry (SpyCas9a) from CasPEDIA.   How can I use it? CasPEDIA...getting the desired features and cutting (note: spCas9 isn’t always the winner!).   Fig. 3 – Comparison...

  16. CRISPR Meets Synthetic Biology: A Conversation with MIT’s Christopher Voigt

    Type
    Blog Post
    ...almost limitless number of DNA binding proteins with Cas9 guided by different promoters. The challenge in ... that stop it from being used. One thing is that Cas9 is toxic. When cells carry CRISPR circuitry, it ...Find the Plasmids: Plasmids for multi-input CRISPR/Cas9 genetic circuits References: Nielsen AA, Voigt ...

  17. CRISPR Plasmids - Bacteria

    Type
    Collection
    ...) Validated In Resistance Co-expressed Cas9 Depositing lab Cas9 species = S. pyogenes (PAM = NGG) 42875... Publication Purify A catalytically inactive Cas9 (dCas9) can be used to purify a region of genomic DNA...immunoprecipitate FLAG-tagged Cas9. Design your gRNA sequence to direct dCas9 to a specific locus, avoiding... pneumoniae Kanamycin none, need Cas9 plasmid Marraffini 42876 pCas9 BsaI E. coli, S. pneumoniae Chloramphenicol...Plasmid Gene/Insert Promoter PI Publication Prime Edit Cas9 H840A nickase fused to a reverse transcriptase (...crRNA. This activity provides a stark contrast to Cas9 and Cpf1, which require that each DNA target have... BBa_J23119 SpeI + HindIII Ampicillin none, need Cas9 plasmid Qi Do you have suggestions for other plasmids...

  18. 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...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...

  19. Genomic Deletions in Mammalian Cell Lines

    Type
    Collection
    ...genomic deletions in mammalian cell lines using CRISPR/Cas9...and Daniel Bauer's labs details the use of CRISPR/Cas9 to create genomic deletions in mammalian cell lines...Genomic Deletions in Mammalian Cell Lines via CRISPR/Cas9. Bauer DE, Canver MC, Orkin SH. J. Vis. Exp. (95...site-specific eukaryotic genome engineering. CRISPR/Cas9 is an inexpensive, facile, and efficient genome ...required DNA yield. Perform maxi-prep for each CRISPR/Cas9 construct. Transfecting CRISPRs into Cells of Interest...NOTE: This protocol involves the delivery of CRISPR/Cas9 plasmids by electroporation 4 . This protocol is...cells. However, transient transfection of CRISPR/Cas9 plasmids may be successfully adapted to numerous...
Showing: 141 - 160 of 314 results