PurposeCRISPR/Cas9 system gRNA cloning
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||61089||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) 2636
Vector typeMammalian Expression, CRISPR ; /Cas9 gRNA cloning vector
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert nameH1 promoter; gRNA sequences
SpeciesH. sapiens (human)
Insert Size (bp)346
- Promoter H1
- Cloning method Restriction Enzyme
- 5′ cloning site EcoRI (not destroyed)
- 3′ cloning site HindIII (not destroyed)
- 5′ sequencing primer M13-F20
- 3′ sequencing primer M13R (Common Sequencing Primers)
As Ampicillin gene and H1 promoter contain BsaI sites, we mutated the sequence of Amp gene (G1601C, not change amino acid), and mutated H1 promoter from GAGACC to GAGGACC to eliminate the BsaI internal site.
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:pUC-H1-gRNA was a gift from Shenglin Huang (Addgene plasmid # 61089 ; http://n2t.net/addgene:61089 ; RRID:Addgene_61089)
For your References section:Precise gene deletion and replacement using the CRISPR/Cas9 system in human cells. Zheng Q, Cai X, Tan MH, Schaffert S, Arnold CP, Gong X, Chen CZ, Huang S. Biotechniques. 2014 Sep 1;57(3):115-24. doi: 10.2144/000114196. eCollection 2014. 10.2144/000114196 PubMed 25209046