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. Author manuscript; available in PMC: 2019 Dec 4.
Published in final edited form as: Cell Rep. 2018 Apr 24;23(4):1220–1229. doi: 10.1016/j.celrep.2018.03.105

Figure 1. Generation and Transformation of Human Cerebral Organoids Using the CRISPR/Cas9 System.

Figure 1

(A) Schematic representation of the CRISPR/Cas9-mediated homologous recombination strategy. CRISPR/Cas9 TP53 sgRNA plasmid was co-electroporated with HRasG12V-expressing homologous recombination donor vector (HRasG12V-tdTomato p53 exon HA), which is also targeted and cleaved by the TP53 sgRNA (orange). Two homologous regions (HA1 and HA2, purple) mediate integration within the TP53 gene locus.

(B) Co-localization of CRISPR/Cas9 (GFP) and HRasG12V (tdTomato) observed 2 weeks after transfection.

(C) Ki-67 immunostaining shows HRasG12V-transduced cells are highly proliferative compared to controls.

(D) Time-lapse imaging of HRasG12V-transduced cells showing invasion from 4 weeks on after electroporation. (a) High-magnification view of the invasive edges of a 13-week transduced organoid.

Scale bars: 100 mm in (B), (C), and (D-a) and 1 mm in (D).