Title
Differential Dna Methylation Drives Allelic Architecture For Grb10-Ddc Locus In The Developing Heart
Date of Award
2021
Degree Type
Dissertation
Degree Name
Doctor of Philosophy (PhD)
Graduate Group
Neuroscience
First Advisor
Marisa S. Bartolomei
Abstract
DNA is organized into spatially distinct units by architectural protein CTCF to enrich for intradomain enhancer-promoter contacts, but developmental and tissue specific regulation of architecture in vivo is poorly defined. By profiling neonatal mouse tissues at the imprinted Grb10-Ddc locus, putative tissue- and allele-specific chromatin was revealed, which we interrogated by deletions in vivo. We uncovered a new tissue-specific intronic insulator at Grb10 that is distinct from the classic differentially methylated imprinting control region, acquiring its own allele-specific DNA methylation during development, ultimately regulating gene expression of the entire cluster in heart and muscle. Termed CTCF-Binding Region 2.3 (CBR2.3), this insulator assembles a paternal-specific contact domain with Ddc located 150 kb away, restricting a newly validated cardiac enhancer to the paternal Ddc promoter. Paternal deletion of CBR2.3 in mice reconfigures the chromatin topology to mimic the maternal chromosome, resulting in Grb10-Ddc misexpression and cardiac phenotypes. This in vivo validation of a robust allelic and tissue-specific insulator offers mechanistic insight into how genes utilize different layers of spatial organization to achieve variation in gene expression in development.
Recommended Citation
Juan, Aimee Marie, "Differential Dna Methylation Drives Allelic Architecture For Grb10-Ddc Locus In The Developing Heart" (2021). Publicly Accessible Penn Dissertations. 5129.
https://repository.upenn.edu/edissertations/5129
Embargoed
Available to all on Friday, January 31, 2025Additional Files
Supp.Table 3.xlsx (5567 kB)Supp.Table 4.xlsx (17 kB)