Lenti-(BB)-EF1a-KRAB-dCas9-P2A-BlastR
(Plasmid #118154)

• Purpose: Catalytically inactive Cas9 from S. pyogenes with P2A-BlastR under the EF1a core promoter, and cloning backbone for sgRNA. Contains BsmBI sites for insertion of spacer sequences.
• Depositing Lab: Jorge Ferrer
• Publication: Ferrer Lab Plasmids (unpublished)

Backbone

• Vector backbone: lentiCRISPRv2
• Backbone manufacturer: Feng Zhang (Addgene #52961)
• Vector type: Mammalian Expression, Lentiviral, CRISPR
• Selectable markers: Blasticidin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: KRAB-dCas9-P2A-BlastR
• Species: H. sapiens (human)
• Insert Size (bp): 4907
• Mutation: See depositor comments below
• Promoter: EF1a core and U6
• Tag / Fusion Protein:
  • HA (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: GATCCGCCACCATGGATGC

Resource Information

• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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

Depositor Comments

We have also generated a version of this backbone with the hPGK promoter driving the expression of KRAB-dCas9. We advise users to test both backbones in their cell type of interest.

Note: Plasmid contains a N1396S mutation in dCas9. This mutation is not known to affect plasmid function.