Figure 5
From: A platform for rapid generation of single and multiplexed reporters in human iPSC lines
Summary of different approaches to generate reporter lines in safe harbors and in endogenous lineage-specific genes.
Genetic modification techniques (ZFNs or TALEN) were used in combination with carefully designed donor vectors in this study to target and modify genes/loci-of-interest in selected parental iPSC lines. The parental iPSC can be well-established control lines, patient-derived lines, pre-engineered lines or master lines for the quick swapping strategy. Depending on the donor vectors and targeting genes/loci, parental iPSC can be engineered or re-engineered into different lines expressing either constitutively active reporter genes at the safe harbors or reporter genes that are in frame downstream of lineage-specific genes. These targeted genetically engineered iPSC can be derived into progenitor cells and further differentiated into different cell types for numerous screening purposes. Additionally, we illustrated a master line cassette exchange strategy which gave us the opportunity to quickly and efficiently generate different reporter lines at the safe harbor sites. Using this strategy, we showed successful targeting to both iPSC and the progenitor cells (solid arrows). Potentially, this strategy can also be applied directly to the differentiated cells (dotted arrow).
