1. Chemical and Biophysical Modulation of Cas9 for Tunable Genome Engineering
- Author
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Lucas B. Harrington, James K. Nuñez, and Jennifer A. Doudna
- Subjects
0301 basic medicine ,CRISPR-Associated Proteins ,Chemical biology ,Computational biology ,Biology ,Biochemistry ,Genome ,Genome engineering ,03 medical and health sciences ,Endonuclease ,0302 clinical medicine ,Genome editing ,Animals ,Guide RNA ,Genetics ,Regulation of gene expression ,Cas9 ,General Medicine ,030104 developmental biology ,Gene Expression Regulation ,biology.protein ,Molecular Medicine ,CRISPR-Cas Systems ,Genetic Engineering ,030217 neurology & neurosurgery ,Protein Binding - Abstract
The application of the CRISPR-Cas9 system for genome engineering has revolutionized the ability to interrogate genomes of mammalian cells. Programming the Cas9 endonuclease to induce DNA breaks at specified sites is achieved by simply modifying the sequence of its cognate guide RNA. Although Cas9-mediated genome editing has been shown to be highly specific, cleavage events at off-target sites have also been reported. Minimizing, and eventually abolishing, unwanted off-target cleavage remains a major goal of the CRISPR-Cas9 technology before its implementation for therapeutic use. Recent efforts have turned to chemical biology and biophysical approaches to engineer inducible genome editing systems for controlling Cas9 activity at the transcriptional and protein levels. Here, we review recent advancements to modulate Cas9-mediated genome editing by engineering split-Cas9 constructs, inteins, small molecules, protein-based dimerizing domains, and light-inducible systems.
- Published
- 2016