Programmable transcriptional repression in mycobacteria using an orthogonal CRISPR interference platform.

ABSTRACT: The development of new drug regimens that allow rapid, sterilizing treatment of tuberculosis has been limited by the complexity and time required for genetic manipulations in Mycobacterium tuberculosis. CRISPR interference (CRISPRi) promises to be a robust, easily engineered and scalable platform for regulated gene silencing. However, in M. tuberculosis, the existing Streptococcus pyogenes Cas9-based CRISPRi system is of limited utility because of relatively poor knockdown efficiency and proteotoxicity. To address these limitations, we screened eleven diverse Cas9 orthologues and identified four that are broadly functional for targeted gene knockdown in mycobacteria. The most efficacious of these proteins, the CRISPR1 Cas9 from Streptococcus thermophilus (dCas9_Sth1), typically a

SUBMITTER: Rock JM 

PROVIDER: S-EPMC5302332 | biostudies-literature | 2017 Feb

REPOSITORIES: biostudies-literature