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. 2014 Jan 20;9(1):e85755. doi: 10.1371/journal.pone.0085755

Figure 3. Modification of the endogenous CCR5 gene by cell-permeable TALEN proteins.

Figure 3

(A) Frequency of endogenous gene disruption in HeLa cells treated with CCR5-targeting TALEN proteins conjugated to R9-CPPs at various (A) peptide-to-protein ratios, (B) TALEN concentrations (µM), and (C) incubation periods (hr). (D) Comparison of gene disruption frequencies in HeLa cells transfected with 200 ng TALEN expression vectors or treated with 1.0 µM TALEN proteins for 2 hr. Gene mutagenesis frequencies were determined by the Surveyor nuclease assay. All R9-conjugated CCR5-targeted TALEN proteins were labeled in the presence of protease inhibitor cocktail. (E) Sequence analysis of the CCR5 locus in HeLa cells treated with 1.0 µM conjugated TALEN proteins for 2 hr. Deletions (dashes) and insertions (lowercase) induced by NHEJ repair are aligned to the wild-type sequence (WT). Black triangles indicate expected Surveyor nuclease assay cleavage products.