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.
| Original language | English |
|---|---|
| Pages (from-to) | 2023-2034 |
| Number of pages | 12 |
| Journal | Biotechnology Letters |
| Volume | 38 |
| Issue number | 12 |
| DOIs | |
| Publication status | Published - 2016 Dec 1 |
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All Science Journal Classification (ASJC) codes
- Biotechnology
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Generation of ΔF508-CFTR T84 cell lines by CRISPR/Cas9-mediated genome editing. / Chung, Woo Young; Song, Myungjae; Park, Jinhong; Namkung, Wan; Lee, Jinu; Kim, Hyongbum; Lee, Min Goo; Kim, Joo Young.
In: Biotechnology Letters, Vol. 38, No. 12, 01.12.2016, p. 2023-2034.Research output: Contribution to journal › Article
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
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 -