UMMS Affiliation
Program in Molecular Medicine; Diabetes Center of Excellence; Graduate School of Biomedical Sciences, Interdisciplinary Graduate Program
Publication Date
2019-04-16
Document Type
Article
Disciplines
Amino Acids, Peptides, and Proteins | Cell Biology | Cells | Developmental Biology | Embryonic Structures | Genetic Phenomena | Nucleic Acids, Nucleotides, and Nucleosides
Abstract
Studies in vertebrates have outlined conserved molecular control of definitive endoderm (END) development. However, recent work also shows that key molecular aspects of human END regulation differ even from rodents. Differentiation of human embryonic stem cells (ESCs) to END offers a tractable system to study the molecular basis of normal and defective human-specific END development. Here, we interrogated dynamics in chromatin accessibility during differentiation of ESCs to END, predicting DNA-binding proteins that may drive this cell fate transition. We then combined single-cell RNA-seq with parallel CRISPR perturbations to comprehensively define the loss-of-function phenotype of those factors in END development. Following a few candidates, we revealed distinct impairments in the differentiation trajectories for mediators of TGFbeta signaling and expose a role for the FOXA2 transcription factor in priming human END competence for human foregut and hepatic END specification. Together, this single-cell functional genomics study provides high-resolution insight on human END development.
Keywords
CRISPRi, chromatin accessibility, dCas9-KRAB, endoderm, hepatic endoderm, human development, perturbation screen, pluripotent stem cells, single-cell RNA-seq, stem cell differentiation
Rights and Permissions
© 2019 The Author(s). This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
DOI of Published Version
10.1016/j.celrep.2019.03.076
Source
Cell Rep. 2019 Apr 16;27(3):708-718.e10. doi: 10.1016/j.celrep.2019.03.076. Link to article on publisher's site
Journal/Book/Conference Title
Cell reports
Related Resources
PubMed ID
30995470
Repository Citation
Genga, Ryan M.; Kernfeld, Eric M.; Parsi, Krishna M.; Parsons, Teagan J.; Ziller, Michael J.; and Maehr, Rene, "Single-Cell RNA-Sequencing-Based CRISPRi Screening Resolves Molecular Drivers of Early Human Endoderm Development" (2019). Open Access Articles. 3818.
https://escholarship.umassmed.edu/oapubs/3818
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.
Included in
Amino Acids, Peptides, and Proteins Commons, Cell Biology Commons, Cells Commons, Developmental Biology Commons, Embryonic Structures Commons, Genetic Phenomena Commons, Nucleic Acids, Nucleotides, and Nucleosides Commons