-
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
-
Depositing Lab
-
Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
---|---|---|---|---|---|
Plasmid | 74630 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
-
Vector backboneAll-in-One_Nickase-D10A
- Backbone size (bp) 9626
-
Vector typeMammalian Expression, CRISPR
- Promoter Cbh
-
Selectable markersPuromycin
-
Tag
/ Fusion Protein
- Puromycin resistant marker
Growth in Bacteria
-
Bacterial Resistance(s)Ampicillin, 100 μg/mL
-
Growth Temperature37°C
-
Growth Strain(s)DH5alpha
-
Copy numberHigh Copy
Cloning Information
- Cloning method Restriction Enzyme
Resource Information
-
Articles Citing this Plasmid
Terms and Licenses
-
Academic/Nonprofit Terms
-
Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
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