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| Status |
Public on Oct 27, 2021 |
| Title |
Single cell RNAseq of WT and BRG1 conditional KO at D4 and D10 of cardiac differentiation |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
We invetigated the transcriptional consequences of BRG1 loss at D4 and D10 stages of cardiac differentiation
Differentiation proceeds along a continuum of increasingly fate-restricted intermediates, referred to as canalization How robust canalization is established or maintained is unclear. Here we show that deletion of the BRG1/BRM-associated factor (BAF) chromatin remodeling complex ATPase gene Brm (encoding Brahma) results in a radical identity switch during directed cardiogenesis of mouse embryonic stem cells (ESCs). Despite establishment of well-differentiated precardiac mesoderm, Brm-null cells subsequently shifted identities, predominantly becoming neural precursors, violating germ layer assignment. Trajectory inference showed sudden acquisition of non-mesodermal identity in Brm-null cells, consistent with a new transition state inducing a saddle-node bifurcation. Mechanistically, loss of Brm prevented de novo accessibility of cardiac enhancers while increasing expression of the neurogenic factor POU3F1 and preventing expression of the neural suppressor REST. Brm mutant identity switch was overcome by increasing BMP4 levels during mesoderm induction, repressing Pou3f1 and re-establishing a cardiogenic chromatin landscape. Our results reveal BRM as a compensable safeguard for fidelity of mesoderm chromatin states, and support a model in which developmental canalization is not a rigid irreversible path, but a highly plastic trajectory that must be safeguarded.
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| Overall design |
single cell RNAseq of WT and BRG1 conditional KO cells during D4 and D10 of cardiac differentiation. The cells were treated with either tetrahydrofuran or with 200nM 4-hydroxytamoxifen in tatrahydrofuran at D2 to have WT and conditional BRG1 KO respectively.
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| Contributor(s) |
Bruneau BG, Hota S |
| Citation(s) |
35082446 |
| |
| Submission date |
May 09, 2020 |
| Last update date |
Feb 04, 2022 |
| Contact name |
Benoit Bruneau |
| E-mail(s) |
[email protected]
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| Organization name |
Gladstone Institute of Cardiovascular Disease
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| Street address |
1650 Owens Street
|
| City |
San Francisco |
| State/province |
CA |
| ZIP/Postal code |
94158 |
| Country |
USA |
| |
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| Platforms (1) |
| GPL19057 |
Illumina NextSeq 500 (Mus musculus) |
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| Samples (4)
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| This SubSeries is part of SuperSeries: |
| GSE150186 |
Brahma safeguards canalization of cardiac mesoderm differentiation |
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| Relations |
| BioProject |
PRJNA631417 |
| SRA |
SRP260913 |
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