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. Author manuscript; available in PMC: 2010 Jul 2.
Published in final edited form as: Cell Stem Cell. 2009 Jun 18;5(1):97–110. doi: 10.1016/j.stem.2009.05.023

Figure 4. HR-mediated gene targeting of EGFP and PIG-A genes in female human MP2 iPS cells.

Figure 4

(A) Microscopy of corrected (GFP+) MP2 iPS cells harboring the EGIP* reporter, 5 days after Nucleofection with GFP-specific ZFNs.

(B) FACS analysis of EGFP gene correction in MP2 iPS cells with or without GFP-specific ZFNs. Dot plots were gated on TRA-1-60+ cells.

(C) PIG-A gene targeting in MP2 iPS cells with or without PIG-A specific ZFNs was performed as described in Figure 3A using the 1:5 ratio of Donor:ZFN DNA. After Nucleofection and hygromycin B selection, TRA-1-60+ (human iPS/ES) cells were analyzed for the presence or absence of CD59 (a GPI-AP). Representative FACS dot plots are shown.

(D) Summary of FACS analysis of CD59- MP2 iPS cell populations after PIG-A gene targeting from 4 independent experiments.

(E) PCR confirms the targeted integration into the PIG-A locus in MP2 iPS cells.