pLenti-dCas9-KRAB-gRNA-TRE-blast
(Plasmid #201152)

• Purpose: Lentiviral expression of S. pyogenes dead Cas9 (dCas9/dSpCas9/SpdCas9) fused to the KRAB domain as well as a gRNA targeting the tight TRE promoter and the blasticidin resistance gene.
• Depositing Lab: Moshe Szyf
• Publication: CRISPRi plasmids for reduced leakiness of doxycycline-inducible tight TRE promoter (unpublished)

Backbone

• Vector backbone: Lenti-(BB)-EF1a-KRAB-dCas9-P2A-BlastR EGFP-guide1
• Backbone manufacturer: Addgene #118158
• Modifications to backbone: Replaced EGFP guide RNA with guide RNA targeting the tight TRE promoter.
• Vector type: Lentiviral, CRISPR
• Selectable markers: Blasticidin

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: dCas9-KRAB
• Alt name: KRAB-dCas9
• Species: Synthetic; S. pyogenes
• Tag / Fusion Protein:
  • HA (C terminal on insert)

Gene/Insert 2

• Gene/Insert name: gRNA-TRE
• Species: Synthetic; S. pyogenes
• Mutation: gRNA sequence: TACGTTCTCTATCACTGAT

Terms and Licenses

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