Gramelspacher, Hou, Zhang (2019) Biochemical characterization of RNA-guided ribonuclease activities for CRISPR-Cas9 systems Methods (San Diego, Calif.) ()

Abstract

The majority of bacteria and archaea rely on CRISPR-Cas systems for RNA-guided, adaptive immunity against mobile genetic elements. The Cas9 family of type II CRISPR-associated DNA endonucleases generates programmable double strand breaks in the CRISPR-complementary DNA targets flanked by the PAM motif. Nowadays, CRISPR-Cas9 provides a set of powerful tools for precise genome manipulation in eukaryotes and prokaryotes. Recently, a few Cas9 orthologs have been reported to possess intrinsic CRISPR-guided, sequence-specific ribonuclease activities. These discoveries fundamentally expanded the targeting capability of CRISPR-Cas9 systems, and promise to provide new CRISPR tools to manipulate specific cellular RNA transcripts. Here we present a detailed method for the biochemical characterization of Cas9's RNA-targeting potential. Copyright © 2019 Elsevier Inc. All rights reserved.

Links

http://www.ncbi.nlm.nih.gov/pubmed/31228550
http://dx.doi.org/10.1016/j.ymeth.2019.06.018

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