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Transcriptional and Post-Transcriptional Polar Effects in Bacterial Gene Deletion Libraries

André Mateus, Malay B. Shah, Johannes F. Hevler, Nils Kurzawa, Jacob Bobonis, Athanasios Typas, Mikhail M. Savitski

2021mSystems33 citationsDOIOpen Access PDF

Abstract

Single-gene deletion libraries have allowed genome-wide characterization of gene function and interactions. While each mutant intends to disrupt the function of a single gene, it can unintentionally target other genes, such as those located in the same operon as the deletion. The extent to which such polar effects occur in deletion libraries has not been assessed. In this work, we use proteomics and transcriptomics data to show that transcript level changes lead to nearly 40% of deletions in operons affecting the protein levels of genes located downstream by at least 2-fold. Furthermore, we observed a post-transcriptional effect on the gene located directly downstream of the deletion. These results can guide the design of future gene deletion libraries and emphasizes the importance of follow-up work when linking genotypes to phenotypes.

Topics & Concepts

OperonGeneBiologyGeneticsTranscriptomeMutantGenomeTranscriptional regulationRegulation of gene expressionGene expressionBacterial Genetics and BiotechnologyRNA and protein synthesis mechanismsCRISPR and Genetic Engineering
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