PurposeU6 driven SaCas9 sgRNA expression vector for cloning own guides
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||122090||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 4286
- Total vector size (bp) 4362
Vector typeMammalian Expression, CRISPR
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert nameSaCas9 tracr
SpeciesSynthetic; S. pyogenes
Insert Size (bp)76
- Promoter U6
- Cloning method Restriction Enzyme
- 5′ cloning site BfuAI (destroyed during cloning)
- 3′ cloning site BfuAI (destroyed during cloning)
- 5′ sequencing primer T3 Promoter
- 3′ sequencing primer T7 Promoter (Common Sequencing Primers)
To design oligos for cloning: 5'-ACCG-Spacer_Sequence-3' and 5'-AAAC-Rev_Comp_Spacer-3'
For sgRNA cloning: Digest with BfuAI and ligate in phosphorylated and annealed oligos
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:SaCas9_sgRNA_expression_in_pBluescript was a gift from Scot Wolfe (Addgene plasmid # 122090 ; http://n2t.net/addgene:122090 ; RRID:Addgene_122090)
For your References section:Orthogonal Cas9-Cas9 chimeras provide a versatile platform for genome editing. Bolukbasi MF, Liu P, Luk K, Kwok SF, Gupta A, Amrani N, Sontheimer EJ, Zhu LJ, Wolfe SA. Nat Commun. 2018 Nov 19;9(1):4856. doi: 10.1038/s41467-018-07310-x. 10.1038/s41467-018-07310-x PubMed 30451839