생화학분자생물학회

	Name: Mahadev Shelake ( rahulms@gnu.ac.kr )
	2019-present	Academic Research Professor, Gyeongsang National University, Jinju
	2017-2019	Senior Researcher, PMBBRC, Gyeongsang National University, Jinju
	2014-2017	Post-doctoral Researcher, Proteo-Science Center, Ehime University, Matsuyama, Japan
	2010-2014	Ph.D., Ehime University, Matsuyama, Japan
	2009-2010	Project Assistant, National Research Center for Grapes, Pune, India
	2008-2009	Research Associate, Mahyco Life Science Research Centre, Jalna, India

	Name: Jae-Yean Kim ( kimjy@gnu.ac.kr )
	2003-present	Professor, Gyeongsang National University, Korea
	2019-2022	President, Korean Society of Plant Breeding Innovation
	2000-2003	Research Fellow, Cold Spring Harbor Laboratory, USA
	1998-2000	Research Fellow, Warwick University, UK

CRISPR base editor-based targeted random mutagenesis (BE-TRM) toolbox for directed evolution.

Directed evolution (DE) of desired locus by targeted random mutagenesis (TRM) tools is a powerful approach for generating genetic variations with novel or improved functions, particularly in complex genomes. TRM-based DE involves developing a mutant library of targeted DNA sequences and screening the variants for the desired properties. However, DE methods have for a long time been confined to bacteria and yeasts. Lately, CRISPR/Cas and DNA deaminase-based tools that circumvent enduring barriers such as longer life cycle, small library sizes, and low mutation rates have been developed to facilitate DE in native genetic environments of multicellular organisms. Notably, deaminase-based base editing-TRM (BE-TRM) tools have greatly expanded the scope and efficiency of DE schemes by enabling base substitutions and randomization of targeted DNA sequences. BE-TRM tools provide a robust platform for the continuous molecular evolution of desired proteins, metabolic pathway engineering, creation of a mutant library of desired locus to evolve novel functions, and other applications, such as predicting mutants conferring antibiotic resistance. This review provides timely updates on the recent advances in BE-TRM tools for DE, their applications in biology, and future directions for further improvements.


BMB Rep. 2023 Dec 6:5950. Online ahead of print.
https://pubmed.ncbi.nlm.nih.gov/38053292/