Myogenic Progenitor Cell Lineage Specification by CRISPR/Cas9-Based Transcriptional Activators.

TitleMyogenic Progenitor Cell Lineage Specification by CRISPR/Cas9-Based Transcriptional Activators.
Publication TypeJournal Article
Year of Publication2020
AuthorsJB Kwon, A Vankara, AR Ettyreddy, JD Bohning, and CA Gersbach
JournalStem Cell Reports
Volume14
Start Page755
Issue5
Pagination755 - 769
Date Published05/2020
Abstract

Engineered CRISPR/Cas9-based transcriptional activators can potently and specifically activate endogenous fate-determining genes to direct differentiation of pluripotent stem cells. Here, we demonstrate that endogenous activation of the PAX7 transcription factor results in stable epigenetic remodeling and differentiates human pluripotent stem cells into skeletal myogenic progenitor cells. Compared with exogenous overexpression of PAX7 cDNA, we find that endogenous activation results in the generation of more proliferative myogenic progenitors that can maintain PAX7 expression over multiple passages in serum-free conditions while preserving the capacity for terminal myogenic differentiation. Transplantation of human myogenic progenitors derived from endogenous activation of PAX7 into immunodeficient mice resulted in a greater number of human dystrophin<sup>+</sup> myofibers compared with exogenous PAX7 overexpression. RNA-sequencing analysis also revealed transcriptome-wide differences between myogenic progenitors generated via CRISPR-based endogenous activation of PAX7 and exogenous PAX7 cDNA overexpression. These studies demonstrate the utility of CRISPR/Cas9-based transcriptional activators for controlling cell-fate decisions.

DOI10.1016/j.stemcr.2020.03.026
Short TitleStem Cell Reports