Figure 3.

Validation of Knockin Cell Lines Generated with CTS

Knockin clones were generated for each disease-associated variant of interest shown in Table S3, with representative clones harboring variants in each gene shown here. Chromatograms (chromats) showing Sanger sequencing results of original cell line (WT) and knockin line (KI) with variants indicated by black arrows. Immunocytochemistry showing pluripotency markers Nanog and SSEA-4 for each knockin cell line harboring the respective variant of interest. Images were merged and counterstained with DAPI. Scale bar, 100 μm. Representative karyotypes for each cell line. Agarose gel showing AAVS1 inside-out PCR for both the 5′ (middle lane) and 3′ (right lane) integration sites (Experimental Procedures), which demonstrates site-specific integration of the selection construct via HDR. See also Figure S3 and Table S3.