Fig. 7
Long-term GESTALT fate mapping of transplanted cells lacking Nanog. (A) Diagram of cell transplantation at sphere stage from a Mnanog; GESTALT barcoded donor embryo into a wild-type host embryo. The donor embryo was injected with sgRNAs targeting CRISPR-Cas9 sites in the GESTALT barcode array. Host animals were grown to 90 days post-fertilization, when intestine, heart, eyes and brain were dissected (n=20 animals across two independent trials). Genomic DNA was prepared and barcodes (corresponding to surviving descendants of Mnanog; GESTALT transplanted cells) were sequenced from each organ. (B) After sequence processing of libraries from each organ across all 20 adults, distinct barcodes corresponding to different clones of transplanted Mnanog; GESTALT cells were counted. Shown is a summary of all 428 clones of Mnanog; GESTALT cells found in host animal organs. |