title: Generation of ΔF508-CFTR T84 cell lines by CRISPR/Cas9-mediated genome editing.
abstract: OBJECTIVE
  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.
authors: Department of Pharmacology and Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, 120-752, Republic of Korea.
authors: Woo Young
authors: WY
authors: Chung
authors: Graduate School of Biomedical Science and Engineering/College of Medicine, Hanyang University, Seoul, 133-791, Republic of Korea.
authors: Myungjae
authors: M
authors: Song
authors: College of Pharmacy, Yonsei Institute of Pharmaceutical Sciences, Yonsei University, Incheon, 406-840, Republic of Korea.
authors: Jinhong
authors: J
authors: Park
authors: College of Pharmacy, Yonsei Institute of Pharmaceutical Sciences, Yonsei University, Incheon, 406-840, Republic of Korea.
authors: Wan
authors: W
authors: Namkung
authors: College of Pharmacy, Yonsei Institute of Pharmaceutical Sciences, Yonsei University, Incheon, 406-840, Republic of Korea.
authors: Jinu
authors: J
authors: Lee
authors: Department of Pharmacology and Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, 120-752, Republic of Korea.
authors: Hyongbum
authors: H
authors: Kim
authors: Department of Pharmacology and Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, 120-752, Republic of Korea.
authors: Min Goo
authors: MG
authors: Lee
authors: Department of Pharmacology and Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, 120-752, Republic of Korea. jooyoungkim@yuhs.ac.
authors: Joo Young
authors: JY
authors: Kim
mesh_terms: D002460:Cell Line; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005434:Flow Cytometry; D000072669:Gene Editing / Q000379:methods*; D006801:Humans; D009154:Mutation
keywords: CRISPR/Cas9; Epithelial cell; Genome editing; T84 cell line; ΔF508-CFTR