Year: 2025
Type: article
Abstract: CRISPR interference (CRISPRi) has emerged as a powerful tool for high-throughput phenotypic screening in bacteria. Its programmability, scalability, and compatibility with non-model bacteria enable systematic genotype–phenotype mapping across diverse biological contexts. This review outlines the current methodologies of CRISPRi-based screening, including sgRNA library construction and screening formats, and highlights a wide range of observable phenotypes—such as gene essentiality, auxotrophy, autotrophy, drug susceptibility, and phage susceptibility. We also discuss recent advancements in strategies for delivering the CRISPRi system to non-model bacteria. Finally, we propose future directions for expanding the utility of CRISPRi-based screening. Together, these insights provide a comprehensive overview of CRISPRi as a versatile platform for bacterial functional genomics and offer perspectives on its continued evolution.
Authors: Jiseon Lee, Jiseon Lee, Jeong Wook Lee, Jeong Wook Lee
Institution: Pohang University of Science and Technology
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $3,890
OpenAlex ID: W4416684883