ATP1A1_G4_Q118R_RUNX1_+4_T_to_G_Dual_pegRNA
(Plasmid #173212)

• Purpose: Control vector for coselection for prime editing in human cells. Tandem expression of ATP1A1 Q118R-G4 and RUNX1 +4 T to G pegRNAs from two independent U6 promoters.
• Depositing Lab: Yannick Doyon
• Publication: Levesque et al Nat Commun. 2022 Oct 7;13(1):5909. doi: 10.1038/s41467-022-33669-z.

Backbone

• Vector backbone: pU6-pegRNA-GG-acceptor
• Total vector size (bp): 2706
• Vector type: Mammalian Expression, CRISPR ; Prime editing

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: ATP1A1 G4 Q118R pegRNA + RUNX1 +4 T to G pegRNA
• gRNA/shRNA sequence: GTTCCTCTTCTGTAGCAGCT and GCATTTTCAGGAGGAAGCGA
• Species: H. sapiens (human)
• Promoter: Tandem U6 promoters

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BsaI (destroyed during cloning)
• 3′ cloning site: BsaI (destroyed during cloning)
• 5′ sequencing primer: CAGGGTTATTGTCTCATGAGCGG
• 3′ sequencing primer: GGGAAACGCCTGGTATCTTT

Resource Information

• Supplemental Documents:
  • ATP1A1_G4_Q118R_RUNX1__4_T_to_G_Dual_pegRNA.gb

Terms and Licenses

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

Depositor Comments

Control pegRNA vector derived from ATP1A1_G4_Q118R_Dual_pegRNA. This vector can be used as a positive control for prime editing (PE3 in combination with ATP1A1_G3_RUNX1_Nick-38_Dual_sgRNA) to co-target ATP1A1 and RUNX1 to perform coselection for RUNX1 +4 T to G. Please visit https://doi.org/10.1101/2021.11.02.464583 for bioRxiv preprint.