Extended Data Fig. 5: CRISPR/Cas9-mediated genome editing of human iPSCs to APOE-KO. | Nature Aging

Extended Data Fig. 5: CRISPR/Cas9-mediated genome editing of human iPSCs to APOE-KO.

From: Proteomics of mouse brain endothelium uncovers dysregulation of vesicular transport pathways during aging

Extended Data Fig. 5

a. APOE knockout strategy: Exon 2 of APOE was targeted by an sgRNA (target and PAM sequence shown), generating a fourteen base pair deletion on one allele and a 13 base pair deletion on the other allele in the APOE KO line. The resulting frameshift exposes a nearby stop codon. b. Immunofluorescence analysis of pluripotency markers, SSEA4, NANOG, TRA160, and OCT 4 with DAPI in APOE KO iPSC line. Scale bar 50 µm. Experiment was repeated 4 times. c. Investigating CRISPR-mediated on-target effects using Sanger sequencing of SNPs near the edited locus in WT and APOE KO iPSC lines showing maintenance of both alleles after editing. d. List of top five most similar off-target sites ranked by the CFD and MIT prediction scores, respectively. Sanger sequencing detected no off-target editing.

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