SP6-hCas9-Ce-mRNA
(Plasmid #47911)

• Purpose: In vitro transcription of a mRNA encoding humanized Cas9 nuclease, with 3' UTRs suitable for germline expression in C. elegans, for doing CRISPR-Cas by RNA injection. Uses SP6 RNA polymerase.
• Depositing Lab: Paul Sternberg
• Publication: Chiu et al Genetics. 2013 Aug 26.

Backbone

• Vector backbone: Bluescript
• Backbone manufacturer: Stratagene
• Backbone size w/o insert (bp): 3000
• Total vector size (bp): 7115
• Vector type: CRISPR

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: hCas9 with C. elegans 5' and 3' UTRs
• Species: H. sapiens (human), C. elegans (nematode)
• Promoter: SP6

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRV (destroyed during cloning)
• 3′ cloning site: KpnI (not destroyed)
• 5′ sequencing primer: T3
• 3′ sequencing primer: T7

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Contains an insert modified from Addgene plasmid #41815 hCas9
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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