PurposeMammalian expression of Cas9 and sgRNA scaffold
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||83480||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Vector backbonelentiCRISPR v2 (#52961)
Backbone manufacturerFeng Zhang
- Backbone size w/o insert (bp) 10000
- Total vector size (bp) 14873
Modifications to backbonepuromycin N-acetyltransferase was replaced by blasticidin S deaminase
Vector typeMammalian Expression, Lentiviral, CRISPR
Growth in Bacteria
Growth Strain(s)NEB Stable
Growth instructionsNo more than 16-hour culture.
Copy numberHigh Copy
Gene/Insert nameBlasticidin S deaminase
MutationReplaced puromycin N-acetyltransferase on the original plasmid
- Promoter EF-1α
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI (not destroyed)
- 3′ cloning site SacII (not destroyed)
- 5′ sequencing primer TGCTGAAACAAGCCGGAGAT
- 3′ sequencing primer AATTAGCCCTTCCAGTCCCC, WPRE_R (Common Sequencing Primers)
lentiCas9-Blast was digested with BamHI and SacII. The resulting small fragment (encoding P2A, blasticidin and a portion of WPRE) was gel-purified. lentiCRISPR v2 was similarly digested and the larger fragment gel-purified. The two fragments combined and ligated.
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:lentiCRISPR v2-Blast was a gift from Mohan Babu (Addgene plasmid # 83480 ; http://n2t.net/addgene:83480 ; RRID:Addgene_83480)