CRISPR-Cas9Optogenetic Tool for Activating Endogenous Genes
Moritoshi Sato's Lab has engineered CRISPR-Cas9 to create a photoactivatable transcription system. The tool allows scientists to use light to spatially and temporally control endogenous gene activation.
This system works by bringing together an anchor protein and activator protein to drive transcription. The anchor consists of dCas9 fused to CIB1, and the anchor is targeted to specific genes via user-defined guide RNAs. Upon stimulation with blue light, the complex recruits a CIB1 binding protein (a portion of CRY2) fused to the p65 activation domain, which activates transcription of downstream genes.
The article describes the targeted activation of endogenous genes and details the time course of gene activation using this system. Scientists can obtain dCas9-trCIB1, CRY2PHR-p65, and other plasmids from this article through Addgene.
· Nihongaki et al., Chemistry and Biology. 2015 Feb 19; 22(2):169-74.