U6-NT sgRNA; EF1a-dCas9-KRAB-GFP
(Plasmid #194284)

• Purpose: EF1a-dCas9-KRAB-GFP with nontarget sgRNA
• Depositing Lab: Elizabeth Kirby
• Publication: Rieskamp et al iScience. 2023 Jun 8;26(7):107068. doi: 10.1016/j.isci.2023.107068. eCollection 2023 Jul 21.

Backbone

• Vector backbone: FUGW; Addgene 71237
• Backbone size w/o insert (bp): 7000
• Total vector size (bp): 14927
• Modifications to backbone: Added NT sgRNA downstream of U6 promoter; replaced hUbC promoter with EF1a
• Vector type: Lentiviral, CRISPR

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

• Gene/Insert name: dCas9-KRAB-GFP
• Alt name: CRSIPRi
• gRNA/shRNA sequence: non target sgRNA
• Species: M. musculus (mouse)
• Promoter: EF1a
• Tag / Fusion Protein:
  • GFP

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Pacl (not destroyed)
• 3′ cloning site: Acc65I (not destroyed)
• 5′ sequencing primer: EF1a-F; U6-F
• 3′ sequencing primer: GFP-R

Resource Information

• Supplemental Documents:
  • p050.ape
• A portion of this plasmid was derived from a plasmid made by: NT sgRNA design from Chiou, S.-H., Winters, I.P., Wang, J., Naranjo, S., Dudgeon, C., Tamburini, F.B., Brady, J.J., Yang, D., Grüner, B.M., Chuang, C.-H., et al. (2015). Pancreatic cancer modeling using retrograde viral vector delivery and in vivo CRISPR/Cas9-mediated somatic genome editing. Genes Dev. 29, 1576–1585. 10.1101/gad.264861.115.

Terms and Licenses

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

Depositor Comments

Please visit https://doi.org/10.1101/2022.10.31.514562 for bioRxiv preprint.