Purpose(Empty Backbone) All-in-One plasmid encoding dual U6 promoter-driven sgRNAs and Cas9-D10A nickase linked via 2A peptide with puromycin resistant marker to enhance efficient and accurate genome editing
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||74630||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
- Backbone size (bp) 9626
Vector typeMammalian Expression, CRISPR
- Promoter Cbh
/ Fusion Protein
- Puromycin resistant marker
Growth in Bacteria
Copy numberHigh Copy
- Cloning method Restriction Enzyme
Terms and Licenses
Puromycin resistance gene is only for transient expression along with Cas9 nickase. Thus, drug selection period should be tested for each experiment. The presence of cellular puromycin resistance could be monitored by performing a control transfection with an empty backbone vector of either AIO-GFP or AIO-mCherry in which GFP/mCherry expression level represents puromycin resistance level.
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:AIO-Puro was a gift from Steve Jackson (Addgene plasmid # 74630 ; http://n2t.net/addgene:74630 ; RRID:Addgene_74630)
For your References section:CRISPR-Cas9(D10A) nickase-based genotypic and phenotypic screening to enhance genome editing. Chiang TW, le Sage C, Larrieu D, Demir M, Jackson SP. Sci Rep. 2016 Apr 15;6:24356. doi: 10.1038/srep24356. 10.1038/srep24356 PubMed 27079678