Kevin R Roy, Justin D Smith, Shengdi Li, Sibylle C Vonesch, Michelle Nguyen, Wallace T Burnett, Kevin M Orsley, Cheng-Sheng Lee, James E Haber, Robert P St Onge, Lars M Steinmetz
Genome editing technologies have the potential to transform our understanding of how genetic variation gives rise to complex traits through the systematic engineering and phenotypic characterization of genetic variants. However, there has yet to be a system with sufficient efficiency, fidelity, and throughput to comprehensively identify causal variants at the genome scale. Here we explored the ability of templated CRISPR editing systems to install natural variants genome-wide in budding yeast. We optimized several approaches to enhance homology-directed repair (HDR) with donor DNA templates, including donor recruitment to target sites, single-stranded donor production by bacterial retrons, and in vivo plasmid assembly...
February 2, 2024: bioRxiv