TY - JOUR
T1 - Generation of ΔF508-CFTR T84 cell lines by CRISPR/Cas9-mediated genome editing
AU - Chung, Woo Young
AU - Song, Myungjae
AU - Park, Jinhong
AU - Namkung, Wan
AU - Lee, Jinu
AU - Kim, Hyongbum
AU - Lee, Min Goo
AU - Kim, Joo Young
N1 - Publisher Copyright:
© 2016, Springer Science+Business Media Dordrecht.
Copyright:
Copyright 2017 Elsevier B.V., All rights reserved.
PY - 2016/12/1
Y1 - 2016/12/1
N2 - 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.
AB - 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.
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U2 - 10.1007/s10529-016-2190-4
DO - 10.1007/s10529-016-2190-4
M3 - Article
C2 - 27571970
AN - SCOPUS:84984653429
VL - 38
SP - 2023
EP - 2034
JO - Biotechnology Letters
JF - Biotechnology Letters
SN - 0141-5492
IS - 12
ER -