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Chromatin and gene regulation during early embryonic development in zebrafish

Hurton, Matthew (2024) Chromatin and gene regulation during early embryonic development in zebrafish. Doctoral Dissertation, University of Pittsburgh. (Unpublished)

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Abstract

Early metazoan embryogenesis features dramatic cellular changes while key regulatory factors act upon the zygotic genome to facilitate its transcriptional competence. This process, called Zygotic Genome Activation (ZGA), is a shift in the transcriptional program, wherein maternal transcription factors engage regulatory sequence of the transcriptionally silent zygotic genome to facilitate a transition from maternal to zygotic control of cellular processes. While much is known about this process in terms of time-scale and factors involved, more precise details of regulatory dynamics remain elusive. In this dissertation I aim to further elucidate the mechanisms by which transcription factors act upon zygotic regulatory sequence to facilitate transcriptional competence and the techniques by which we can probe this phenomenon. I have done so in primarily three ways. First, I have adapted a ChIP-Seq alternative, CUT&RUN, for early zebrafish embryos, and used it to assay a wide variety of histone modifications both known to be and putatively associated with active regulatory sequence to develop a richer understanding of the regulatory environment during ZGA. In doing so, I discovered novel regulatory sequences that may represent a more ancient regulatory paradigm for early embryonic transcriptional regulation. Second, I have tested CRISPR mediated loss-of-function techniques on zebrafish embryos, as well as generated several loss-of-function mutant lines for key maternal regulatory factors. Third, I worked toward adapting CUT&Tag to design a strategy to measure bivalent chromatin at regulatory sequence. Together, this thesis further characterized early zygotic gene regulation by both providing novel techniques and resources for further study and identifying previously unknown regulatory mechanisms that are important for early zygotic transcription.

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Details

Item Type: University of Pittsburgh ETD
Status: Unpublished
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Hurton, Matthewmdh84@pitt.edumdh840009-0001-7842-8985
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Committee ChairLee, Milermiler@pitt.edumlee0000-0003-0933-0551
Committee MemberArndt, Karenarndt@pitt.edukarndt0000-0003-1320-9957
Committee MemberCarlson, Anneacarlson@pitt.eduacarlson0000-0003-2724-1325
Committee MemberRebeiz, Markrebeiz@pitt.edumrebeiz0000-0001-5731-5570
Committee MemberTsang, Michaeltsang@pitt.edumtsang0000-0001-7123-0063
Date: 13 May 2024
Date Type: Publication
Defense Date: 27 March 2024
Approval Date: 13 May 2024
Submission Date: 4 April 2024
Access Restriction: 2 year -- Restrict access to University of Pittsburgh for a period of 2 years.
Number of Pages: 157
Institution: University of Pittsburgh
Schools and Programs: Dietrich School of Arts and Sciences > Biological Sciences
Degree: PhD - Doctor of Philosophy
Thesis Type: Doctoral Dissertation
Refereed: Yes
Uncontrolled Keywords: developmental biology; transcriptionalal regulation; embryogenesis;
Date Deposited: 13 May 2024 13:49
Last Modified: 13 May 2024 13:49
URI: http://d-scholarship.pitt.edu/id/eprint/46013

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