생화학분자생물학회

	Name: Junho Hur ( juhur@hanyang.ac.kr )
	2020-present	Associate Professor, Department of Genetics, College of Medicine, Hanyang University
	2017-2020	Assistant Professor, Department of Pathology, College of Medicine, Kyung Hee University
	2016-2017	Research Professor, Korea University
	2015-2016	Research Associate, Institute for Basic Science
	2012-2015	Postdoctoral Fellow, California Institute of Technology, USA
	2006-2012	Ph.D., Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, USA

Development of CRIPSR technology for precise single-base genome editing: a brief review

Clustered regularly interspaced short palindromic repeats (CRISPR) was originally found as a type of bacterial immune system. In many CRISPR systems, the functional ribonucleoproteins (RNPs) are composed of CRISPR protein and guide RNAs, and they selectively bind and cleave specific target DNAs or RNAs, based on the sequence complementarity with the guide RNA. The specific targeted cleavage of the nucleic acids by CRISPR has been broadly utilized in genome editing methods. In the process of genome editing of eukaryotic cells, CRISPR-mediated DNA double strand breaks (DSB) at specific genomic loci activate the endogenous DNA repair systems and induce mutations at the target sites with high efficiencies. Two of the major endogenous DNA repair machineries are non-homologous end joining (NHEJ) and homology-directed repair (HDR). In the event of DSB, the two repair pathways operate in competition, resulting in several possible outcomes including deletions, insertions, and substitutions. Due to the inherent stochasticity of DSB-based genome editing methods, it was difficult to achieve defined single-base changes without unanticipated random mutation patterns. In order to overcome the heterogeneity issue in DSB-mediated genome editing, novel methods have been recently developed to install precise single-base level changes without inducing DSB. The line of approaches utilized catalytically compromised CRISPR in conjunction with base-modifying enzymes and DNA polymerases, to accomplish highly efficient and precise genome editing of single and multiple bases. In this review, we introduce some of the advances in the single-base level CRISPR genome editing methods and their applications.


https://pubmed.ncbi.nlm.nih.gov/33298245/
BMB Rep. 2021 Feb;54(2):98-105.