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pCAG-denAsCas12a(E174R/S542R/K548R/D908A)-NLS(nuc)-3xHA-VPR (RTW776)
(Plasmid #107943)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 107943 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pCAG-CFP
  • Backbone manufacturer
    Connie Cepko lab (Addgene plasmid #11179)
  • Vector type
    Mammalian Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    human codon optimized DNase-inactive (D908A) enAsCas12a (E174R/S542R/K548R) fused to VPR activation domain
  • Alt name
    RTW776
  • Alt name
    inactive version of enhanced Cas12a from Acidaminococcus sp. BV3L6
  • Species
    Acidaminococcus sp. BV3L6
  • Mutation
    E174R, S542R, K548R and D908A
  • Promoter CAG
  • Tags / Fusion Proteins
    • NLS (nucleoplasmin) (C terminal on insert)
    • 3x HA (C terminal on insert)
    • VPR (VP64-p65-Rta) (C terminal on insert)

Cloning Information

  • Cloning method Gibson Cloning
  • 5′ sequencing primer pCAG-F (5'-GCAACGTGCTGGTTATTGTG-3')
  • 3′ sequencing primer Bglob-pA-R (5'-ttttggcagagggaaaaaga-3')
    • (Common Sequencing Primers)

Resource Information

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.
How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pCAG-denAsCas12a(E174R/S542R/K548R/D908A)-NLS(nuc)-3xHA-VPR (RTW776) was a gift from Keith Joung & Benjamin Kleinstiver (Addgene plasmid # 107943 ; http://n2t.net/addgene:107943 ; RRID:Addgene_107943)

  • For your References section:

    Engineered CRISPR-Cas12a variants with increased activities and improved targeting ranges for gene, epigenetic and base editing. Kleinstiver BP, Sousa AA, Walton RT, Tak YE, Hsu JY, Clement K, Welch MM, Horng JE, Malagon-Lopez J, Scarfo I, Maus MV, Pinello L, Aryee MJ, Joung JK. Nat Biotechnol. 2019 Feb 11. pii: 10.1038/s41587-018-0011-0. doi: 10.1038/s41587-018-0011-0. 10.1038/s41587-018-0011-0 PubMed 30742127
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