AI Article Synopsis
- Understanding the function of bacterial genes is challenging, but double-mutant genetic interaction analysis helps identify how genes work together by linking unknown genes to established pathways.
- The research introduces double-CRISPRi as a method to measure genetic interactions on a large scale, even for essential genes, leading to the discovery of over 1000 known and new interactions.
- Findings highlight the unique roles of similar genes and uncover new genes tied to cell division, showcasing double-CRISPRi's potential for exploring bacterial gene networks in future studies.
Article Abstract
Understanding bacterial gene function remains a major biological challenge. Double-mutant genetic interaction (GI) analysis addresses this challenge by uncovering the functional partners of targeted genes, allowing us to associate genes of unknown function with novel pathways and unravel connections between well-studied pathways, but is difficult to implement at the genome-scale. Here, we develop and use double-CRISPRi to systematically quantify genetic interactions at scale in the envelope, including essential genes. We discover > 1000 known and novel genetic interactions. Our analysis pipeline and experimental follow-ups reveal the distinct roles of paralogous genes such as the and actin homologs, and identify new genes involved in the well-studied process of cell division. Overall, our study provides valuable insights into gene function and demonstrates the utility of double-CRISPRi for high-throughput dissection of bacterial gene networks, providing a blueprint for future studies in diverse bacterial species.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11343205 | PMC |
| http://dx.doi.org/10.1101/2024.08.14.608006 | DOI Listing |
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