PurposeConstitutive expression of Cas9 gene and inducible expression of the plasmid curing system for CRISPR mediated genome editing of E. coli.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||71538||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Vector typeBacterial Expression, CRISPR
Growth in Bacteria
Gene/Insert namePlasmid curing system
Alt namegRNA targeting bla gene
- Promoter pBAD
Cloning Information for Gene/Insert 2
- Cloning method Unknown
- 5′ sequencing primer unknown (Common Sequencing Primers)
The plasmid curing system consists of an arabinose inducible promoter, PBAD to express a gRNA targeting the bla gene on the gRNA plasmid. When induced, the bla-targeting gRNA is expressed, leading Cas9 to cleave the gRNA plasmid and resulting in plasmid elimination.
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:pCas9cur was a gift from Tao Chen (Addgene plasmid # 71538 ; http://n2t.net/addgene:71538 ; RRID:Addgene_71538)
For your References section:Metabolic engineering of Escherichia coli using CRISPR-Cas9 meditated genome editing. Li Y, Lin Z, Huang C, Zhang Y, Wang Z, Tang YJ, Chen T, Zhao X. Metab Eng. 2015 Sep;31:13-21. doi: 10.1016/j.ymben.2015.06.006. Epub 2015 Jun 30. 10.1016/j.ymben.2015.06.006 PubMed 26141150