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VVT1
(Plasmid #65779)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 65779 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    MLM3636
  • Backbone manufacturer
    Joung lab (Addgene Plasmid # 43860)
  • Vector type
    Mammalian Expression, CRISPR

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    XL1 Blue
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    SaCas9 gRNA backbone, without spacer sequence
  • Species
    Synthetic
  • Promoter U6

Cloning Information

Resource Information

Depositor Comments

Use BsmBI sites to insert sgRNA spacer sequence into the vector. See the supplemental document "Joung Lab gRNA Cloning Protocol" for more details.

UPDATE 5/31/2016: The Joung Lab recommends using BPK2660 (Addgene plasmid # 70709) instead of VVT1 as an SaCas9 guide RNA cloning vector, as guide RNAs cloned into BPK2660 are more effective than those cloned into VVT1. The difference in efficacy is due to VVT1 containing the full length guide RNA (predicted from the natural CRISPR sequence in S. aureus), whereas BPK2660 uses a truncated gRNA that is more effective. For the same oligo/spacer, the rate of mutagenesis is greater using BPK2660. See Kleinstiver et al., 2015 Nat Biotech for additional information on the truncated gRNA plasmid (https://www.ncbi.nlm.nih.gov/pubmed/26524662).

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:

    VVT1 was a gift from Keith Joung (Addgene plasmid # 65779 ; http://n2t.net/addgene:65779 ; RRID:Addgene_65779)
  • For your References section:

    Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Kleinstiver BP, Prew MS, Tsai SQ, Topkar VV, Nguyen NT, Zheng Z, Gonzales AP, Li Z, Peterson RT, Yeh JJ, Aryee MJ, Joung JK. Nature. 2015 Jun 22. doi: 10.1038/nature14592. 10.1038/nature14592 PubMed 26098369