pLV hU6-sgRNA hUbC-dCas9-KRAB-T2a-Puro
(Plasmid #71236)

• Purpose: Express sgRNA and dCas9-KRAB from lentiviral vector
• Depositing Lab: Charles Gersbach
• Publication: Thakore et al Nat Methods. 2015 Dec;12(12):1143-9. doi: 10.1038/nmeth.3630. Epub 2015 Oct 26.

Backbone

• Vector backbone: FUGW
• Total vector size (bp): 15000
• Vector type: Lentiviral, CRISPR
• Selectable markers: Puromycin

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: humanized dCas9-KRAB T2A Puro
• Species: S. Pyogenes
• Mutation: D10A and H840A
• Tag / Fusion Protein:
  • Flag (N terminal on insert)

Gene/Insert 2

• Gene/Insert name: sgRNA

Resource Information

• Addgene Notes:
  • Addgene Diagnostic Digest- MluI
  • Addgene Diagnostic Digest- NotI/KpnI
• Articles Citing this Plasmid:
  • 93 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Note: DNA prep concentrations may be low due to the size of this plasmid, so working with low copy protocols, higher starting volumes, or fresh transformants is recommended.