Optimizing sgRNA structure to improve CRISPR-Cas9 knockout efficiency.

ABSTRACT:

Background

Single-guide RNA (sgRNA) is one of the two key components of the clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 genome-editing system. The current commonly used sgRNA structure has a shortened duplex compared with the native bacterial CRISPR RNA (crRNA)-transactivating crRNA (tracrRNA) duplex and contains a continuous sequence of thymines, which is the pause signal for RNA polymerase III and thus could potentially reduce transcription efficiency.

Results

Here, we systematically investigate the effect of these two elements on knockout efficiency and showed that modifying the sgRNA structure by extending the duplex length and mutating the fourth thymine of the continuous sequence of thymines to cytosine or guanine significantly, and sometime

SUBMITTER: Dang Y 

PROVIDER: S-EPMC4699467 | biostudies-literature | 2015 Dec

REPOSITORIES: biostudies-literature