AM77_pU6.Sa-gRNA.CLYBL
(Plasmid #199237)

• Purpose: Expression construct encoding a S. aureus guide RNA targeting the human CLYBL safe harbor locus
• Depositing Lab: Manuel Goncalves
• Publication: Wang et al Nucleic Acids Res. 2023 Mar 17:gkad165. doi: 10.1093/nar/gkad165.

Backbone

• Vector backbone: pBR322
• Total vector size (bp): 2290
• Vector type: Mammalian Expression, CRISPR, Synthetic Biology

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: S. aureus gRNA spacer
• Species: H. sapiens (human), Synthetic
• Insert Size (bp): 23
• Promoter: RNA polymerase III promoter for human U6 snRNA

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BsmBI (destroyed during cloning)
• 3′ cloning site: BsmBI (destroyed during cloning)
• 5′ sequencing primer: M13 rev
• 3′ sequencing primer: M13 fwd

Resource Information

• Supplemental Documents:
  • AM77_pU6.Sa-gRNA.CLYBL.gbk

Terms and Licenses

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

Depositor Comments

This gRNA expression construct can be combined with target site-containing donor plasmids AD59_pEP.DonorCLYBL.TS (Addgene plasmid #199227) or AZ64_pE.DonorCLYBL.TS (Addgene plasmid #199228) to insert foreign DNA at the human CLYBL safe harbor locus using SaCas9 nucleases (HMEJ-based gene targeting) or SaCas9.D10A nickases (ITPN-based gene targeting), such as that encoded by Addgene plasmid #199251.