PurposeFor in vivo expression of Cas9 or generation of Cas9 mRNA
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||47322||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 4095
Vector typeMammalian Expression, CRISPR ; xenopus and zebrafish expression
Growth in Bacteria
Copy numberHigh Copy
Insert Size (bp)4100
- Promoter SP6
- Cloning method Restriction Enzyme
- 5′ cloning site ClaI (not destroyed)
- 3′ cloning site XbaI (not destroyed)
- 5′ sequencing primer SP6
- 3′ sequencing primer M13R (Common Sequencing Primers)
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.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pCS2-Cas9 was a gift from Alex Schier (Addgene plasmid # 47322 ; http://n2t.net/addgene:47322 ; RRID:Addgene_47322)
For your References section:Efficient mutagenesis by Cas9 protein-mediated oligonucleotide insertion and large-scale assessment of single-guide RNAs. Gagnon JA, Valen E, Thyme SB, Huang P, Ahkmetova L, Pauli A, Montague TG, Zimmerman S, Richter C, Schier AF. PLoS One. 2014 May 29;9(5):e98186. doi: 10.1371/journal.pone.0098186. eCollection 2014. PONE-D-14-14210 [pii] PubMed 24873830