High-Throughput Gene Mutagenesis Screening Using Base Editing

Abstract

Base editing is a CRISPR-Cas9 genome engineering tool that allows programmable mutagenesis without the creation of double-stranded breaks. Here, we describe the design and execution of large-scale base editing screens using the Target-AID base editor in yeast. Using this approach, thousands of sites can be mutated simultaneously. The effects of these mutations on fitness can be measured using a pooled growth competition assay followed by DNA sequencing of gRNAs as barcodes.

Keywords: Base editing; CRISPR-Cas9; Functional genomics; High-throughput; Mutagenesis; Systems biology; Target-AID; Yeast.