A universal method for sensitive and cell-free detection of CRISPR-associated nucleases.

ABSTRACT: A multitude of biological applications for CRISPR-associated (Cas) nucleases have propelled the development of robust cell-based methods for quantitation of on- and off-target activities of these nucleases. However, emerging applications of these nucleases require cell-free methods that are simple, sensitive, cost effective, high throughput, multiplexable, and generalizable to all classes of Cas nucleases. Current methods for cell-free detection are cumbersome, expensive, or require sophisticated sequencing technologies, hindering their widespread application beyond the field of life sciences. Developing such cell-free assays is challenging for multiple reasons, including that Cas nucleases are single-turnover enzymes that must be present in large excess over their substrate and that diffe

SUBMITTER: Cox KJ 

PROVIDER: S-EPMC6419926 | biostudies-literature | 2019 Mar

REPOSITORIES: biostudies-literature