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The position of the target site for engineered nucleases improves the aberrant mRNA clearance in in vivo genome editing

Authors
 Jae Hoon Lee  ;  Sungsook Yu  ;  Tae Wook Nam  ;  Jae-il Roh  ;  Young Jin  ;  Jeong Pil Han  ;  Ji-Young Cha  ;  Yoon Ki Kim  ;  Su-Cheong Yeom  ;  Ki Taek Nam  ;  Han-Woong Lee 
Citation
 SCIENTIFIC REPORTS, Vol.10(1) : e4173, 2020 
Journal Title
 SCIENTIFIC REPORTS 
Issue Date
2020
Abstract
Engineered nucleases are widely used for creating frameshift or nonsense mutations in the target genes to eliminate gene functions. The resulting mRNAs carrying premature termination codons can be eliminated by nonsense-mediated mRNA decay. However, it is unclear how effective this process would be in vivo. Here, we found that the nonsense-mediated decay was unable to remove the mutant mRNAs in twelve out of sixteen homozygous mutant mice with frameshift mutations generated using engineered nucleases, which is far beyond what we expected. The frameshift mutant proteins translated by a single nucleotide deletion within the coding region were also detected in the p53 mutant mice. Furthermore, we showed that targeting the exons present downstream of the exons with a start codon or distant from ATG is relatively effective for eliminating mutant mRNAs in vivo, whereas the exons with a start codon are targeted to express the mutant mRNAs. Of the sixteen mutant mice generated, only four mutant mice targeting the downstream exons exhibited over 80% clearance of mutant mRNAs. Since the abnormal products, either mutant RNAs or mutant proteins, expressed by the target alleles might obscure the outcome of genome editing, these findings will provide insights in the improved performance of engineered nucleases when they are applied in vivo.
Files in This Item:
T202000826.pdf Download
DOI
10.1038/s41598-020-61154-4
Appears in Collections:
1. College of Medicine (의과대학) > BioMedical Science Institute (의생명과학부) > 1. Journal Papers
Yonsei Authors
Nam, Ki Taek(남기택)
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/175615
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