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 |
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Pages (from-to) | 2023-2034 |
Number of pages | 12 |
Journal | Biotechnology Letters |
Volume | 38 |
Issue number | 12 |
DOIs | |
Publication status | Published - 2016 Dec 1 |
Bibliographical note
Funding Information:This work was supported by NRF-2011-002945, NRF-2011-0023701, and 2013R1A3A2042197 from the National Research Foundation (NRF), the Ministry of Science, ICT & Future Planning, Korea.
Publisher Copyright:
© 2016, Springer Science+Business Media Dordrecht.
All Science Journal Classification (ASJC) codes
- Biotechnology
- Bioengineering
- Applied Microbiology and Biotechnology