pCS2-Cas9
(Plasmid #47322)

• Purpose: For in vivo expression of Cas9 or generation of Cas9 mRNA
• Depositing Lab: Alex Schier
• Publication: Gagnon et al PLoS One. 2014 May 29;9(5):e98186. doi: 10.1371/journal.pone.0098186. eCollection 2014.

Backbone

• Vector backbone: pCS2
• Backbone size w/o insert (bp): 4095
• Vector type: Mammalian Expression, CRISPR ; xenopus and zebrafish expression

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: Cas9
• Insert Size (bp): 4100
• Promoter: SP6

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: ClaI (not destroyed)
• 3′ cloning site: XbaI (not destroyed)
• 5′ sequencing primer: SP6
• 3′ sequencing primer: M13R

Resource Information

• Articles Citing this Plasmid:
  • 10 References

Terms and Licenses

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

Depositor Comments

Please see the Schier lab website http://labs.mcb.harvard.edu/schier/ for detailed protocols about target site selection, sgRNA production, stop codon cassette design, Cas9 protein purification, injection and downstream analysis.

This plasmid was used to prepare and inject Cas9 mRNA into zebrafish zygotes.