Figure 1 | Scientific Reports

Figure 1

From: CRISPR/Cas9-mediated generation of a tyrosine hydroxylase reporter iPSC line for live imaging and isolation of dopaminergic neurons

Figure 1

Generation of the TH-mOrange reporter iPSC line using CRISPR/Cas9-mediated gene editing. (A) Scheme describing the recombination steps during the edition process. Blue arrows represent the primers used for the PCR screening procedure. Black triangles represent LoxP sites surrounding the selection cassette. (B) Molecular analysis of the correctly targeted clones to confirm proper P2A-mOrange cassette integration and selection cassette excision in the control iPSC line. Full-length gels are included in Fig. S1C. (C) Sanger sequencing confirmed successful excision of the LoxP site-flanked cassette. (D) Immunofluorescence analysis of representative colonies of the TH reporter (SP_11) iPSC line staining positive for the pluripotency-associated markers NANOG, OCT4 and SOX2 (green) and TRA-1-81, SSEA3 and SSEA4 (red). Scale bar, 50 µm. (E) Normal karyotype of the TH reporter control iPSC line.

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