Purpose(Empty Backbone) Human expression plasmid for SaCas9 sgRNA(84nt in length) (need to clone in spacer into BsmBI sites): U6-BsmBIcassette-Sa-sgRNA(84)
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||70709||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Backbone manufacturerJoachim Messing (Addgene plasmid # 50005)
Vector typeMammalian Expression, CRISPR
- Promoter U6
Growth in Bacteria
Copy numberHigh Copy
- Cloning method Restriction Enzyme
- 5′ sequencing primer OS280 (5'-CAGGGTTATTGTCTCATGAGCGG-3') (Common Sequencing Primers)
Use BsmBI sites to insert sgRNA spacer sequence.
The length of the sgRNA (not including the spacer sequence) is 84 nucleotides, where this truncated repeat/anti-repeat sgRNA is similar to that described previously by Ran et al., Nature 2015 (PubMed ID: 25830891) and conveys higher activity than the canonical full length sgRNA
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:BPK2660 was a gift from Keith Joung (Addgene plasmid # 70709 ; http://n2t.net/addgene:70709 ; RRID:Addgene_70709)
For your References section:Broadening the targeting range of Staphylococcus aureus CRISPR-Cas9 by modifying PAM recognition. Kleinstiver BP, Prew MS, Tsai SQ, Nguyen NT, Topkar VV, Zheng Z, Joung JK. Nat Biotechnol. 2015 Nov 2. doi: 10.1038/nbt.3404. 10.1038/nbt.3404 PubMed 26524662