Systematic identification and characterization of high efficiency Cas9 guide RNAs for therapeutic targeting of ADAR.

ABSTRACT: Therapeutic targeting of the adenosine deaminase ADAR has great potential in cancer and other indications; however, it remains unclear what approach can enable effective and selective therapeutic inhibition. Herein, we conduct multi-staged guide RNA screening and identify high efficiency Cas9 guide RNAs to enable a CRISPR/Cas-based approach for ADAR knockout. Through characterization in human primary immune cell systems we observe similar activity with two-part guide RNA and single guide RNA, dose responsive activity, similar guide activity rank order across different cell types, and favorable computational off-target profiles of candidate guide RNAs. We determine that knockout of ADAR using these guide RNAs induces pharmacodynamic responses primarily consisting of immunological responses

SUBMITTER: Gowen BG 

PROVIDER: S-EPMC11849909 | biostudies-literature | 2025

REPOSITORIES: biostudies-literature