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CRISPR base editor-based targeted random mutagenesis (BE-TRM) toolbox for directed evolutionopen access

Authors
Shelake, Rahul MahadevPramanik, DibyajyotiKim, Jae-Yean
Issue Date
Jan-2024
Publisher
The Biochemical Society of the Republic of Korea
Keywords
Base editing; CRISPR/Cas; Directed evolution; Dual base editors; Targeted random mutagenesis
Citation
BMB reports, v.57, no.1, pp 30 - 39
Pages
10
Indexed
SCIE
SCOPUS
KCI
Journal Title
BMB reports
Volume
57
Number
1
Start Page
30
End Page
39
URI
https://scholarworks.gnu.ac.kr/handle/sw.gnu/69685
DOI
10.5483/bmbrep.2023-0086
ISSN
1976-6696
1976-670X
Abstract
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 Reports 2024; 57(1): 30-39].
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