Identification of Cis-Regulatory elements specific for the Neural Crest during Xenopus development

Hyde, Anne Cathrine (2022) Identification of Cis-Regulatory elements specific for the Neural Crest during Xenopus development. Masters thesis, University of East Anglia.

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The neural crest (NC) is a multipotent cell population that develops early in the embryo. The NC cells (NCC) migrate throughout the embryo and contribute to a range of different tissues including the craniofacial skeleton, the heart, and many different neurons. To achieve this multipotency, the NC is highly regulated by cis-regulatory elements (CREs) which can decide the special and temporal expression of genes required for NC formation. Some CREs specific for NC have been identified in a several different species, but few CREs specific for NC development have been identified in Xenopus.

CREs are believed to exist in open regions of the genome. Here we use an assay for transposase accessible chromatin using sequencing (ATAC-seq) to identify these open regions in Xenopus animal cap tissue induced to be NC. We have identified putative CREs from this data using bioinformatic approaches and then tested the activity of these putative CREs in a reporter assay by creating transgenic Xenopus embryos using a I-Scel meganuclease method.

We initially identified and cloned 20 putative CREs, and of these 12 showed either neural or specific NC developmental expression and are therefore believed to regulate different genes previously associated with NC development. These include putative CREs for sox9 and snai2.

The regulation of NC development is very complex and to understand diseases associated with NC we need to understand this regulation. We have identified a good method for cloning the putative CREs identified in ATAC-seq data into a reporter vector for creating transgenic Xenopus embryos. This method has allowed us to identify several putative CREs specific for NC development.

Item Type: Thesis (Masters)
Faculty \ School: Faculty of Science > School of Biological Sciences
Depositing User: Chris White
Date Deposited: 28 Apr 2022 13:50
Last Modified: 28 Apr 2022 13:56


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