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Targeted Nucleotide Editing Technologies for Microbial Metabolic Engineering
Author(s) -
Arazoe Takayuki,
Kondo Akihiko,
Nishida Keiji
Publication year - 2018
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.201700596
Subject(s) - crispr , genome editing , recombineering , biology , genome engineering , computational biology , genome , guide rna , gene , cas9 , genetics
Since the emergence of programmable RNA‐guided nucleases based on clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR‐associated protein (Cas) systems, genome editing technologies have become a simplified and versatile tool for genome editing in various organisms and cell types. Although genome editing enables efficient genome manipulations, such as gene disruptions, gene knockins, and chromosomal translocations via DNA double‐strand break (DSB) repair in eukaryotes, DSBs induced by the CRISPR/Cas system are lethal or severely toxic to many microorganisms. Therefore, in many prokaryotes, including industrially useful microbes, the CRISPR/Cas system is often used as a negative selection component in combination with recombineering or other related strategies. Novel and revolutionary technologies have been recently developed to re‐write targeted nucleotides (C:G to T:A and A:T to G:C) without DSBs and donor DNA templates. These technologies rely on the recruitment of deaminases at specific target loci using the nuclease‐deficient CRISPR/Cas system. Here, the authors review and compare CRISPR‐based genome editing, current base editing platforms and their spectra. The authors discuss how these technologies can be applied in various aspects of microbial metabolic engineering to overcome barriers to cellular regulation in prokaryotes.