Skip to main content
. 2014 Nov 3;3(12):1418–1428. doi: 10.5966/sctm.2014-0102

Figure 3.

Figure 3.

Lineage-specific reporter line generation by zinc-finger nuclease (ZFN)-mediated gene targeting. The process for creating knock-in induced pluripotent stem cell (iPSC) lines by ZFN technology is illustrated. ZFN mRNAs and donor vector are inserted into the iPSC via nucleofection, and iPSCs that were successfully integrated were selected for via drug selection. An example of a donor vector with a reporter and a selection marker strategy is shown. The surviving clones (∼10–50 clones picked) are expanded for 2-4 weeks, with genomic DNA used to screen and confirm for mutations. The targeted iPSC lines are then assayed for pluripotency and genomic stability and other characterization desired. Abbreviations: 2A, 2A peptide; d, day; gDNA, genomic DNA; GFAP, Glial fibrillary acidic protein; LA, left homologous recombination arm; Neo, Neomycin resistant gene; PGK, phosphoglycerate kinase promoter; Puro, puromycin resistant gene; RA, right homologous recombination arm.