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Microbial Base Editing: A Powerful Emerging Technology for Microbial Genome Engineering
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Microbial Base Editing: A Powerful Emerging Technology for Microbial Genome Engineering
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Microbial Base Editing: A Powerful Emerging Technology for Microbial Genome Engineering
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Journal Article
Microbial Base Editing: A Powerful Emerging Technology for Microbial Genome Engineering
2021
Overview
Genome engineering is crucial for answering fundamental questions about, and exploring practical applications of, microorganisms. Various microbial genome-engineering tools, including CRISPR/Cas-enhanced homologous recombination (HR), have been developed, with ever-improving simplicity, efficiency, and applicability. Recently, a powerful emerging technology based on CRISPR/Cas-nucleobase deaminase fusions, known as base editing, opened new avenues for microbial genome engineering. Base editing enables nucleotide transition without inducing lethal double-stranded (ds)DNA cleavage, adding foreign donor DNA, or depending on inefficient HR. Here, we review ongoing efforts to develop and apply base editing to engineer industrially and clinically relevant microorganisms. We also summarize bioinformatics tools that would greatly facilitate guide (g)RNA design and sequencing data analysis and discuss the future challenges and prospects associated with this technology.
Base editing represents an emerging genome-editing technology that cleverly combines the programmability of CRISPR/Cas and the catalytic activity of nucleobase deaminase, enabling point mutations at multiple target loci without generating double-strand DNA breaks, requiring exogenous DNA donors, or relying on homology-directed repair.Simplicity, accuracy, and multiplex editing capability facilitate the quick adoption of base editing in the genetic manipulation of many industrially and clinically relevant microorganisms.User-friendly bioinformatics tools have been developed to design base editing-tailored guide RNAs and quantify editing efficiency from sequencing data.Developments in engineering DNA binding and deamination modules for expanded genome-targeting scope and adjustable editing window render base editing a more powerful microbial genome-engineering technology.
