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Generation of ΔF508-CFTR T84 cell lines by CRISPR/Cas9-mediated genome editing

Authors
 Woo Young Chung  ;  Myungjae Song  ;  Jinhong Park  ;  Wan Namkung  ;  Jinu Lee  ;  Hyongbum Kim  ;  Min Goo Lee  ;  Joo Young Kim 
Citation
 Biotechnology Letters, Vol.38(12) : 2023-2034, 2016 
Journal Title
 Biotechnology Letters 
ISSN
 0141-5492 
Issue Date
2016
MeSH
Cell Line ; Clustered Regularly Interspaced Short Palindromic Repeats/genetics ; Endoplasmic Reticulum/metabolism ; Flow Cytometry ; Gene Editing/methods* ; Humans ; Mutation
Keywords
CRISPR/Cas9 ; Epithelial cell ; Genome editing ; T84 cell line ; ΔF508-CFTR
Abstract
OBJECTIVES: To provide a simple method to make a stable ΔF508-CFTR-expressing T84 cell line that can be used as an efficient screening model system for ΔF508-CFTR rescue. RESULTS: CFTR knockout cell lines were generated by Cas9 with a single-guide RNA (sgRNA) targeting exon 1 of the CFTR genome, which produced indels that abolished CFTR protein expressions. Next, stable ΔF508-CFTR expression was achieved by genome integration of ΔF508-CFTR via the lentivirus infection system. Finally, we showed functional rescue of ΔF508-CFTR not only by growing the cells at a low temperature, but also incubating with VX-809, a ΔF508-CFTR corrector, in the established T84 cells expressing ΔF508-CFTR. CONCLUSIONS: This cell system provides an appropriate screening platform for rescue of ΔF508-CFTR, especially related to protein folding, escaped from endoplasmic-reticulum-associated protein degradation, and membrane transport.
Full Text
https://link.springer.com/article/10.1007%2Fs10529-016-2190-4
DOI
10.1007/s10529-016-2190-4
Appears in Collections:
1. Journal Papers (연구논문) > 1. College of Medicine (의과대학) > Dept. of Pharmacology (약리학교실)
Yonsei Authors
김주영(Kim, Joo Young) ORCID logo https://orcid.org/0000-0003-2623-1491
김형범(Kim, Hyongbum) ORCID logo https://orcid.org/0000-0002-4693-738X
이민구(Lee, Min Goo) ORCID logo https://orcid.org/0000-0001-7436-012X
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URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/152015
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