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.
journal_name
Biotechnol Lettjournal_title
Biotechnology lettersauthors
Chung WY,Song M,Park J,Namkung W,Lee J,Kim H,Lee MG,Kim JYdoi
10.1007/s10529-016-2190-4subject
Has Abstractpub_date
2016-12-01 00:00:00pages
2023-2034issue
12eissn
0141-5492issn
1573-6776pii
10.1007/s10529-016-2190-4journal_volume
38pub_type
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