Tetsushi Sakuma

TL;DR: To restore the dystrophin protein, three correction methods were performed in DMD-patient-derived iPSCs, and exon knockin was the most effective approach and provides an important framework for developing iPSC-based gene therapy for genetic disorders using programmable nucleases.

TL;DR: It is demonstrated that CRISPR/Cas9-mediated PITCh, termed CRIS-PITCh, can be applied in human cells without carrying the plasmid backbone sequence and will be useful for a variety of applications, not only in cultured cells, but also in various organisms, including invertebrates and vertebrates.

TL;DR: It is shown that human induced pluripotent stem cells differentiate into incipient mesoderm-like cells (iMeLCs), which robustly generate human primordial germ cell- like cells (hPGCLCs) that can be purified using the surface markers EpCAM and INTEGRINα6.

TL;DR: This work established a system for constructing all-in-one expression vectors containing multiple guide RNA expression cassettes and a Cas9 nuclease/nickase expression cassette that provides an efficient targeting strategy for multiplex genome/epigenome editing, simultaneous activation/repression of multiple genes, and beyond.

TL;DR: A streamlined protocol for PITCh knock-in is described, including the design and construction of the PITCh vectors, and their delivery to either human cell lines by transfection or to frog embryos by microinjection.