pPB_mU6_enh-gRNA_Puro-T2A-BFP
(Plasmid #235571)

• Purpose: Enhanced guide RNA expression & genomic integration. Target gRNA sequences are cloned in via the BstXI and BlpI sites.
• Depositing Lab: Jamie Hackett
• Publication: Policarpi et al Nat Genet. 2024 Jun;56(6):1168-1180. doi: 10.1038/s41588-024-01706-w. Epub 2024 May 9.

Backbone

• Vector backbone: pPB-U6
• Vector type: CRISPR

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: sgRNA backbone
• gRNA/shRNA sequence: Unspecified
• Species: Synthetic

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Unknown (unknown if destroyed)

Resource Information

• Supplemental Documents:
  • pPB_mU6_enh-gRNA_Puro-T2A-BFP.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Evrogen Limited Use Label License for FPs
  • piggyBac Limited Use Label License
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Drives expression of a sub-cloned single guide RNA (gRNA) with enhanced scaffold to target epigenetic editing to a desired locus. gRNA are cloned using the BlpI and BstXI sites. This is a piggyBAC plasmid that integrates into the genome and can be selected for via Puromycin resistance. An additional tagBFP insert is expressed constituitively and indirectly indicates presence of the gRNA.