- 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;
- Collins, James J;
- Lewis, Nathan E;
- Kiani, Samira;
- Church, George M
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.