1. An enhanced CRISPR repressor for targeted mammalian gene regulation
- Author
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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, and Church, George M
- Subjects
Biochemistry and Cell Biology ,Biological Sciences ,Biotechnology ,Genetics ,Human Genome ,CRISPR-Associated Protein 9 ,CRISPR-Cas Systems ,Gene Expression Regulation ,Gene Silencing ,Genes ,Synthetic ,HEK293 Cells ,Humans ,Methyl-CpG-Binding Protein 2 ,RNA ,Guide ,Kinetoplastida ,Recombinant Fusion Proteins ,Repressor Proteins ,Technology ,Medical and Health Sciences ,Developmental Biology ,Biological sciences - 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.
- Published
- 2018