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BmB Reports

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

  • 작성자

    Mahadev Shelake
  • 작성일자

    2023-12-21
  • 조회수

    1832
Name: Mahadev Shelake ( rahulms@gnu.ac.kr )
2019-presentAcademic Research Professor, Gyeongsang National University, Jinju
2017-2019Senior Researcher, PMBBRC, Gyeongsang National University, Jinju
2014-2017Post-doctoral Researcher, Proteo-Science Center, Ehime University, Matsuyama, Japan
2010-2014Ph.D., Ehime University, Matsuyama, Japan
2009-2010Project Assistant, National Research Center for Grapes, Pune, India
2008-2009Research Associate, Mahyco Life Science Research Centre, Jalna, India
Name: Jae-Yean Kim ( kimjy@gnu.ac.kr )
2003-present Professor, Gyeongsang National University, Korea
2019-2022President, Korean Society of Plant Breeding Innovation
2000-2003Research Fellow, Cold Spring Harbor Laboratory, USA
1998-2000Research 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/