Fig. 6

Construction and function of Slc26a3 P131R genetic variant on murine colonic epithelial (CMT-93) cells. a Generation of murine CMT-93 cells carrying the P131R SNP using the clustered regularly interspaced short palindromic repeats/Cas9 system. Outline of the targeting strategy to generate knock-in cell lines using a pair of composite Cas9n–sgRNA expression vectors and a donor single-stranded DNA oligonucleotide (ssODN) targeting the wild-type allele c in the Slc26a3 gene. b Results of Sanger sequencing for confirming the construction of P131R SNP on CMT-93 cells by CRISPR/Cas9. c CMT-93 cells were seeded and cultured overnight in growth medium, the monolayers were starved from serum for 2 h, then treated with 150 mM of NaCl. Representative figure of trans epithelial electric resistance (TEER) data from ECIS analysis of wild-type CMT-93 cells, Caco-2 cells containing the rs386833481(c.392C>G; p.P131R) SNP generated by CRISPR/Cas9 knock-in CMT-93 cells. d P131R Slc26a3 in CMT-93 cells significantly decreased the 150 mM NaCl-induced decrease in TEER value compared with WT CMT-93 cells. *P < 0.05, **P < 0.01, ***P < 0.001