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. 2020 Feb 18;1(3):203–215. doi: 10.34067/KID.0000422019

Figure 1.

Figure 1.

Engineering APOL1 G1 kidney organoids from induced pluripotent stem cells. (A) Schematic summarizing the CRISPR-Cas9 approach to knocking in the G1 risk variants rs73885319 and rs60910145 into the 1016SevA line, including the chosen protospacer, homology-directed repair (HDR) donor template design leveraging the piggyBac transposon system, and Sanger sequencing validation of successful variant knock-in and selection cassette excision. (B) Overview of the Freedman kidney organoid differentiation protocol, with light microscopy of the 1016SevA line forming tubular organoids. (C) Confocal immunofluorescence images of nephron markers in representative G0 and G1 organoids on day 21 of differentiation. E-Cad, E-cadherin; gDNA, genomic DNA; mTeSR1, Modified Tenneille’s Special Recipe 1.