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CRISPR/Cas9 recombineering-mediated deep mutational scanning of essential genes in Escherichia coli.

Molecular systems biology - 1 Mar 2020

Choudhury Alaksh, Fenster Jacob A, Fankhauser Reilly G, Kaar Joel L, Tenaillon Olivier, Gill Ryan T

Abstract excerpt

Deep mutational scanning can provide significant insights into the function of essential genes in bacteria. Here, we developed a high-throughput method for mutating essential genes of Escherichia coli in their native genetic context. We used Cas9-mediated recombineering to introduce a library of mutations, created by error-prone PCR, within a gene fragment on the genome using a single gRNA pre-validated for high...

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Identifiers

PMID: 32175691
DOI: 10.15252/msb.20199265
PMCID: PMC7073797

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