Yonsei Advanced Science Institute

Here, we found two genomic safe harbor (GSH) candidates from chromosome 3 and 8, based on a large-scale population-based cohort data from 4,694 Koreans by CNV analysis. Furthermore, estimated genotype of these CNVRs was validated by quantitative real-time PCR (qRT-PCR), and epidemiological data examined no significant genetic association between diseases/traits and two CNVRs. After screening the GSH candidates by in silico approaches, we designed TALEN pairs to integrate eGFP expression cassette into human cell lines in order to confirm the functionality of GSH candidates at in vitro setting. As a result, transgene insertion into one of the two loci using TALEN showed robust transgene expression comparable to that into AAVS1 site without significantly perturbing neighboring genes. Changing the promoter or cell type did not noticeably disturb this trend. Thus, we could validate two CNVRs as a site for effective and safe transgene insertion in human cells.