Evaluation of CERT gene disruption by TALEN-CERT pairs in HeLa cells.

A, Target sites of TALENs-CERT pairs (version 1 and 2) in human CERT gene. The sequences are located in exon 2, which codes part of the PH domain. The target sites are shown in bold. The numbers on the right and left of the sequence indicate the sequence numbers from the A of the translation initiation codon, based on CERT mRNA (accession number AY4533859). B, Resistance to lysenin in TALEN-CERT–treated HeLa cells. Six TALEN-CERT pairs, which contain two pairs of repeat modules directed against different target sites (CERTv1 and v2) contained in three TALEN scaffolds (TAL original (ori), TAL-ModA, and TAL-ModB), as well as the empty vector (–). These TALEN pairs were transfected into HeLa-mCAT#8 cells, followed by 30°C incubation. The cells were treated with lysenin at 800 ng/ml for 2 h. Their viability was estimated by the MTT assay and is expressing as a percentage of the value (OD570) in the absence of lysenin: mean percentage ± S.D. obtained from three independently repeated experiments. The Bonferroni corrected t-test was used for multiple comparisons. *, p