pLenti CMV GFP Neo (Addgene Plasmid #17447) was used for the construction of expression vectors. DNA fragments encoding mouse claudin-3, claudin-3 lacking the COOH terminus KDYV (delYV), and claudin-3 carrying C113S, C116S, C192S, and C194S mutations (4S) were subcloned into the pLenti CMV GFP Neo vector. Claudin-null cells were generated by using the CRSPR-Cpf1 system. The target sequences were as follows:
claudin-1 (mouse), 5′-ATCCTGGCTTCTCTGGGATGGAT-3′; claudin-3 (mouse), 5′-GGCCTTCATCGGCAGCAGCATCA-3′; claudin-4 (mouse), 5′-CCTCTTCTGCCCGGAAGCCACCA-3′; claudin-7 (mouse), 5′-GCAGCTCATCATGCCGGTGCTCT-3′; claudin-9 (mouse), 5′-GTGTCCTGTGCCCTGCCACTGTG-3′; and claudin-10b (mouse), 5′-GCCAACCTGTGGAAGATCTGCGT-3′.
claudin-1 (mouse), 5′-ATCCTGGCTTCTCTGGGATGGAT-3′; claudin-3 (mouse), 5′-GGCCTTCATCGGCAGCAGCATCA-3′; claudin-4 (mouse), 5′-CCTCTTCTGCCCGGAAGCCACCA-3′; claudin-7 (mouse), 5′-GCAGCTCATCATGCCGGTGCTCT-3′; claudin-9 (mouse), 5′-GTGTCCTGTGCCCTGCCACTGTG-3′; and claudin-10b (mouse), 5′-GCCAACCTGTGGAAGATCTGCGT-3′.
