Light-controlled genome editing by activation of Cas9-mRNA translation.

ABSTRACT: Genome editing by the nuclease Cas9 and guide RNAs enables precise inactivation of genes and presents the basis for numerous research tools and emerging therapies. A critical aspect is the nuclease activity causing off-target effects. Approaches to control where and when active Cas9 is present are therefore desirable. Using Cas9-mRNA already presents a viable way to limit nuclease activity temporally but does not permit controlled induction. Here, we show that Cas9 activity is readily obtained by irradiation of cells transfected with a translationally muted Cas9-mRNA. Using a dual reporter system, we confirm light-mediated knockout of the eGFP-gene by flow cytometry, fluorescence microscopy, Western blotting and sequencing. This system does not involve photocaged proteins nor photocaged gu

SUBMITTER: Schepers H 

PROVIDER: S-EPMC12230757 | biostudies-literature | 2025 Jul

REPOSITORIES: biostudies-literature