2024/05/01 by Yuki Noguchi, Risa Matsui, J. P. Suh +2 · 1 voice
Biochemistry, Genetics and Molecular Biology · #Animal Genetics and Reproduction #CRISPR and Genetic Engineering #Viral Infectious Diseases and Gene Expression in Insects
paper · doi:10.1146/annurev-genom-121222-115958
openalex publication_date 2024/05/01 · openalex created_date 2025/10/10 · openalex updated_date 2026/07/22
Genome-wide screening is a potent approach for comprehensively understanding the molecular mechanisms of biological phenomena. However, despite its widespread use in the past decades across various biological targets, its application to biochemical reactions with temporal and reversible biological outputs remains a formidable challenge. To uncover the molecular machinery underlying various biochemical reactions, we have recently developed the revival screening method, which combines flow cytometry-based cell sorting with library reconstruction from collected cells. Our refinements to the traditional genome-wide screening technique have proven successful in revealing the molecular machinery of biochemical reactions of interest. In this article, we elucidate the technical basis of revival screening, focusing on its application to CRISPR-Cas9 single guide RNA (sgRNA) library screening. Finally, we also discuss the future of genome-wide screening while describing recent achievements from in vitro and in vivo screening.