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In vivo gene correction with targeted sequence substitution through microhomology-mediated end joining

Authors
 Jeong Hong Shin  ;  Soobin Jung  ;  Suresh Ramakrishna  ;  Hyongbum Henry Kim  ;  Junwon Lee 
Citation
 Biochemical and Biophysical Research Communications, Vol.502(1) : 116-122, 2018 
Journal Title
 Biochemical and Biophysical Research Communications 
ISSN
 0006-291X 
Issue Date
2018
MeSH
Animals ; DNA End-Joining Repair* ; Disease Models, Animal ; Genetic Therapy/methods* ; HEK293 Cells ; Humans ; Hydrolases/genetics* ; Mice ; Mutation ; RNA, Guide/genetics ; RNA, Messenger/genetics ; Tyrosinemias/genetics* ; Tyrosinemias/pathology ; Tyrosinemias/therapy*
Keywords
Hereditary tyrosinemia ; Homology-directed repair (HDR) ; In vivo gene correction ; Microhomology-mediated end-joining (MMEJ) ; Targeted sequence substitution (TSS)
Abstract
Genome editing technology using programmable nucleases has rapidly evolved in recent years. The primary mechanism to achieve precise integration of a transgene is mainly based on homology-directed repair (HDR). However, an HDR-based genome-editing approach is less efficient than non-homologous end-joining (NHEJ). Recently, a microhomology-mediated end-joining (MMEJ)-based transgene integration approach was developed, showing feasibility both in vitro and in vivo. We expanded this method to achieve targeted sequence substitution (TSS) of mutated sequences with normal sequences using double-guide RNAs (gRNAs), and a donor template flanking the microhomologies and target sequence of the gRNAs in vitro and in vivo. Our method could realize more efficient sequence substitution than the HDR-based method in vitro using a reporter cell line, and led to the survival of a hereditary tyrosinemia mouse model in vivo. The proposed MMEJ-based TSS approach could provide a novel therapeutic strategy, in addition to HDR, to achieve gene correction from a mutated sequence to a normal sequence.
Full Text
https://www.sciencedirect.com/science/article/pii/S0006291X18311999
DOI
10.1016/j.bbrc.2018.05.130
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Pharmacology (약리학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Ophthalmology (안과학교실) > 1. Journal Papers
Yonsei Authors
Kim, Hyongbum(김형범) ORCID logo https://orcid.org/0000-0002-4693-738X
Lee, Jun Won(이준원) ORCID logo https://orcid.org/0000-0003-0543-7132
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/165509
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