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

ABSTRACT: Myogenic Progenitor Cell Lineage Specification by CRISPR/Cas9-based Transcriptional Activators

PROVIDER: PRJNA607567 | ENA |

REPOSITORIES: ENA

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

Project description: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 directly reprograms human pluripotent stem cells into skeletal myoblast progenitor cells. Compared to the 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+ myofibers compared to exogenous PAX7 overexpression. RNA-seq 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 progenitor cell specification and their potential for regenerative medicine.

2020-04-10 | GSE145575 | GEO

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

Project description: Not available

| S-EPMC7221109 | biostudies-literature

Comparative Analysis of Cas9 Activators Across Multiple Species

Project description:RNA-Seq after Cas9-gRNA transfection with different activators targeting HBG1

2016-04-26 | GSE80611 | GEO

Temporal mechanisms of myogenic specification in human induced pluripotent stem cells

Project description:Understanding the mechanisms of myogenesis in human induced pluripotent stem cells (hiPSCs) is a prerequisite to achieving patient-specific therapy for diseases of skeletal muscle. hiPSCs of different origin show distinctive kinetics and ability to differentiate into myocytes. To address the unique cellular and temporal context of hiPSC differentiation, we perform a longitudinal comparison of the transcriptomic profiles of three hiPSC lines that display differential myogenic specification, one robust and two blunted. We detail temporal differences in mechanisms that lead to robust myogenic specification. We show gene expression signatures of putative cell subpopulations and extracellular matrix components that may support myogenesis. Further, we show that targeted knockdown of ZIC3 at the outset of differentiation leads to improved myogenic specification in blunted hiPSC lines. Our study suggests that β-catenin transcriptional cofactors mediate crosstalk between multiple cellular processes and exogenous cues to facilitate specification of hiPSCs to mesoderm lineage, leading to robust myogenesis.

2021-03-01 | GSE161025 | GEO

Myogenic progenitor specification from pluripotent stem cells.

Project description: Not available

| S-EPMC5723218 | biostudies-other

Muscle progenitor specification and myogenic differentiation are associated with changes in chromatin topology.

Project description:Muscle progenitor specification and myogenic differentiation are associated with changes in chromatin topology.

| PRJNA633082 | ENA

Temporal mechanisms of myogenic specification in human induced pluripotent stem cells

Project description:Temporal mechanisms of myogenic specification in human induced pluripotent stem cells

| PRJNA675129 | ENA

CRISPR-Cas9 RBM10 synthetic lethality screen

Project description:Genome-wide CRISPR-Cas9 knockout screen using TKOv1 sgRNA library was performed in isogenic RBM10-proficient and RBM10-deficient HCC827 cells.

2023-03-04 | E-MTAB-12724 | biostudies-arrayexpress

Uncovering mechanisms of early human lineage specification by CRISPR/Cas9-mediated genome editing

Project description:Uncovering mechanisms of early human lineage specification by CRISPR/Cas9-mediated genome editing

| PRJNA392661 | ENA

Muscle progenitor specification and myogenic differentiation are associated with changes in chromatin topology

Project description:Using Hi-C, promoter-capture Hi-C (pCHi-C), and other genome-wide approaches in skeletal muscle progenitors that inducibly express a master transcription factor, Pax7, we systematically characterized at high-resolution the spatio-temporal re-organization of compartments and promoter-anchored interactions as a consequence of myogenic commitment and differentiation. To further investigate transcriptional regulatory mechanisms governed by Pax7, we used immuno-affinity purification and mass spectrometric sequencing to identify the compendium of Pax7-associated proteins in muscle progenitors. Although Pax7-associated proteins have been identified in myoblasts, it is likely that such an approach would fail to uncover progenitor specific interactions with this protein. We therefore isolated chromatin from iPax7 cells engineered with a single, inducible copy of the Flag-tagged Pax7 transgene integrated next to the Hprt locus, which is expected to maintain expression levels comparable to those of satellite cells in the presence of Dox. Through purification of Flag-Pax7, we identified a cohort of factors and complexes involved in gene activation or repression and remodeling of chromatin and genome architecture that were substantially enriched compared to the uninduced control. We also identified a large cohort of sequence-specific TFs, including several that were shown to play an essential role in muscle stem cells (e.g., Foxk1, Six1, Runx1, Tead1, Nfix) and that are recruited to Pax7-bound enhancers in muscle cells. Importantly, we also confirmed multiple interactions from our proteomic screen through immunoprecipitation and western blotting.

2020-10-30 | MSV000086392 | MassIVE

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