pAAV_LP1B_St1Cas9_LMD-9_SpA_U6_sgRNA
(Plasmid #110624)

• Purpose: A single vector AAV-Cas9 system containing a liver-specific promoter with Cas9 from Streptococcus thermophilus CRISPR1 (St1Cas9 LMD-9) and its U6-driven sgRNA
• Depositing Lab: Yannick Doyon
• Publication: Agudelo et al Genome Res. 2020 Jan;30(1):107-117. doi: 10.1101/gr.255414.119. Epub 2020 Jan 3.

Backbone

• Vector backbone: pX602 (Plasmid #61593)
• Total vector size (bp): 6743
• Modifications to backbone: BsmBI sites near Amp cassette removed
• Vector type: Mammalian Expression, Mouse Targeting, AAV, 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 1

• Gene/Insert name: St1Cas9 LMD-9
• Species: H. sapiens (human), Synthetic
• Promoter: LP1B
• Tags / Fusion Proteins:
  • SV40 NLS (N terminal on insert)
  • SV40 NLS (C terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: U6

Gene/Insert 2

• Gene/Insert name: sgRNA for St1Cas9
• Species: H. sapiens (human), Synthetic
• Promoter: hU6

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: U6

Resource Information

• Supplemental Documents:
  • pAAV_LP1B_St1Cas9_LMD-9_SpA_U6_sgRNA.gb
  • Plasmid Description and Cloning Protocol

Terms and Licenses

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

Depositor Comments

Mammalian codon-optimized Streptococcus thermophilus CRISPR1-Cas9 LMD-9 was subcloned from MSP1594 (Plasmid #65775). An SV40 NLS was added to the N-terminus. Please visit https://www.biorxiv.org/content/early/2018/05/14/321208 for BioRxiv preprint