Anti-CRISPR-mediated continuous directed evolution of CRISPR-Cas9 in human cells.

ABSTRACT: Engineering CRISPR-Cas systems for improved or altered function is central to both research and therapeutic applications. Unfortunately most optimization, especially directed evolution in bacterial hosts, fails to capture the functional requirements of the complex mammalian cellular milieu, where activity is usually required. Robust strategies to enable continuous directed evolution of genome-targeting agents directly in human cells remain lacking. Here, we introduce CRISPR-MACE (Mammalian cell-enabled Adenovirus-assisted Continuous Evolution) as a foundational technology to address this need. CRISPR-MACE integrates virus-based continuous evolution with anti-CRISPR-based tunable selection to generate novel Streptococcus pyogenes Cas9 variants with both increased and decreased DNA bi

SUBMITTER: Sabol AL 

PROVIDER: S-EPMC12710652 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature