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. 2017 Jun 30;9(9):1224–1243. doi: 10.15252/emmm.201607137

Figure EV5. CRISPR/Cas9 editing of CFTR alleles to generate an HBE cell line constitutively expressing ΔI1234_R1239‐CFTR.

Figure EV5

  1. Engineering strategy to delete 18 nt (red) in both alleles of the CFTR gene of HBE cells expressing WT‐CFTR.
  2. Sequencing confirms deletion of 18 nt and presence of engineered splice site donor, C>G (gray box). The arrow indicates the boundary between adjacent exons.
  3. Immunoblots of HBE cell lines endogenously expressing WT‐CFTR, ΔF508‐CFTR and ΔI1234_R1239‐CFTR (left panel), as well as comparison to an HBE cell line in which CFTR alleles were disrupted (i.e., ‘knocked out’, HBE‐KO). Band B, black arrowhead; band C, white arrowhead; *, non‐specific mAb artifact.