Combining CRISPRi and metabolomics for functional annotation of compound libraries.

ABSTRACT: Molecular profiling of small molecules offers invaluable insights into the function of compounds and allows for hypothesis generation about small-molecule direct targets and secondary effects. However, current profiling methods are limited in either the number of measurable parameters or throughput. Here we developed a multiplexed, unbiased framework that, by linking genetic to drug-induced changes in nearly a thousand metabolites, allows for high-throughput functional annotation of compound libraries in Escherichia coli. First, we generated a reference map of metabolic changes from CRISPR interference (CRISPRi) with 352 genes in all major essential biological processes. Next, on the basis of the comparison of genetic changes with 1,342 drug-induced metabolic changes, we made de novo predi

SUBMITTER: Anglada-Girotto M 

PROVIDER: S-EPMC7612681 | biostudies-literature | 2022 May

REPOSITORIES: biostudies-literature