1. An enhanced CRISPR repressor for targeted mammalian gene regulation
- Author
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Massachusetts Institute of Technology. Department of Biological Engineering, Collins, James J, Collins, James J., Yeo, Nan Cher, Chavez, Alejandro, Lance-Byrne, Alissa, Chan, Yingleong, Menn, David, Milanova, Denitsa, Kuo, Chih-Chung, Guo, Xiaoge, Sharma, Sumana, Tung, Angela, Cecchi, Ryan J., Tuttle, Marcelle, Pradhan, Swechchha, Lim, Elaine T., Davidsohn, Noah, Ebrahimkhani, Mo R., Lewis, Nathan E., Kiani, Samira, Church, George M., Massachusetts Institute of Technology. Department of Biological Engineering, Collins, James J, Collins, James J., Yeo, Nan Cher, Chavez, Alejandro, Lance-Byrne, Alissa, Chan, Yingleong, Menn, David, Milanova, Denitsa, Kuo, Chih-Chung, Guo, Xiaoge, Sharma, Sumana, Tung, Angela, Cecchi, Ryan J., Tuttle, Marcelle, Pradhan, Swechchha, Lim, Elaine T., Davidsohn, Noah, Ebrahimkhani, Mo R., Lewis, Nathan E., Kiani, Samira, and Church, George M.
- Abstract
The RNA-guided endonuclease Cas9 can be converted into a programmable transcriptional repressor, but inefficiencies in target-gene silencing have limited its utility. Here we describe an improved Cas9 repressor based on the C-terminal fusion of a rationally designed bipartite repressor domain, KRAB–MeCP2, to nuclease-dead Cas9. We demonstrate the system’s superiority in silencing coding and noncoding genes, simultaneously repressing a series of target genes, improving the results of single and dual guide RNA library screens, and enabling new architectures of synthetic genetic circuits., Paul G. Allen Frontiers Group
- Published
- 2019