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Single-cell RNA Sequencing and Analysis of Human Pancreatic Islets
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Single-Cell RNA-Sequencing-Based CRISPRi Screening Resolves Molecular Drivers of Early Human Endoderm Development
Ryan M J Genga1, Eric M Kernfeld1, Krishna M Parsi1
1Program in Molecular Medicine, Diabetes Center of Excellence, University of Massachusetts Medical School, Worcester, MA 01605, USA.
Cell Reports
|April 18, 2019
Summary
Human embryonic stem cell (ESC) differentiation to definitive endoderm (END) reveals human-specific molecular regulation. This study identifies key transcription factors, including FOXA2, crucial for human END development and foregut/hepatic specification.
Area of Science:
- Developmental Biology
- Stem Cell Biology
- Genomics
Background:
- Vertebrate definitive endoderm (END) development is molecularly conserved.
- Human END development exhibits unique regulatory mechanisms compared to rodents.
- Human embryonic stem cells (ESCs) provide a model for studying human END development.
Purpose of the Study:
- To investigate chromatin accessibility dynamics during human ESC to END differentiation.
- To identify DNA-binding proteins regulating human END cell fate transitions.
- To functionally characterize candidate factors using single-cell RNA-seq and CRISPR perturbations.
Main Methods:
- Interrogation of chromatin accessibility dynamics during ESC differentiation.
- Prediction of DNA-binding proteins involved in END development.
- Single-cell RNA sequencing combined with parallel CRISPR screening for functional genomics.
Main Results:
- Identified key regulators of human END development.
- Revealed distinct differentiation impairments for TGFβ signaling mediators.
- Demonstrated FOXA2's role in priming human END for foregut and hepatic specification.
Conclusions:
- This study provides high-resolution insight into human END development.
- Functional genomics approach elucidates human-specific molecular mechanisms.
- FOXA2 is critical for establishing human END competence for specific organ lineages.
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