pEJS-HZ79 All-in-one AAV-U1a-NmeABE8e-2xBPSV40-U6-Rosa26_V2
(Plasmid #199261)

• Purpose: Optimized single AAV vector for expressing N-terminal fusion Nme2Cas9-ABE8e and one U6 driven sgRNA targeting mouse Rosa26 gene. This optimized construct showed improved in vivo editing efficiency.
• Depositing Lab: Erik Sontheimer
• Publication: Zhang et al GEN Biotechnol. 2022 Jun 1;1(3):285-299. doi: 10.1089/genbio.2022.0015. Epub 2022 Jun 14.

Backbone

• Vector backbone: AAV
• Backbone size w/o insert (bp): 3486
• Total vector size (bp): 7482
• Modifications to backbone: Contains U6-driven sgRNA cassette
• Vector type: Mammalian Expression, AAV, CRISPR

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: NmeABE8e
• Alt name: Nme2Cas9-ABE8e
• Alt name: Nme2Cas9c-ABE8e
• Species: N. meningitidis
• Insert Size (bp): 3996
• Promoter: U1a promoter
• Tags / Fusion Proteins:
  • NLS (N terminal on insert)
  • NLS (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: gccaccatgaaaagaaccgc
• 3′ sequencing primer: ctagagcgcttacacaaaaaacca

Resource Information

• Supplemental Documents:
  • pEJS-HZ79 All-in-one AAV-U1a-NmeABE8e-2xBPSV40-U6-Rosa26_V2.gb

Terms and Licenses

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