A functional screen of novel haematopoietic players in haematopoietic ontogeny

Tvardauskaite, Melita (2024). A functional screen of novel haematopoietic players in haematopoietic ontogeny. University of Birmingham. M.Sc.

Preview

Tvardauskaite2024MScbyRes.pdf
Text - Accepted Version
Available under License All rights reserved.
Download (2MB) | Preview

Abstract

All blood cells in the adult differentiate from a common precursor, the haematopoietic stem and progenitor cell (HSPC), which arise from the haemogenic endothelium (HE) in the dorsal aorta, migrate to an intermediary haematopoietic organ (CHT in zebrafish) to proliferate late seed the definitive haematopoietic organs. Many genetic and epigenetic mechanisms regulate these processes; however many questions remain unanswered regarding the exact mechanisms of this regulation. By analysing recently published data from haematopoietic tissues including the HE and emerging HSPCs, this thesis aimed to identify novel players in HSPC ontogeny, and functionally analyse them. Seven candidate genes were selected, and functionally analysed using CRISPR-cas9. Loss of sart3, wasla, ptpn13, tead1a, esrp1, etv5b, and spi2 all resulted in impaired haematopoietic development. Performing differential gene expression analysis, genes upregulated in the HE were identified, and their functional co-expression network constructed. This revealed a further two genes which are co-expressed together with spi2 – thy1 and ncf1. Upon knockout both genes showed haematopoietic phenotypes in terms of HSPC and lineage marker expression. Further analysis is needed to characterise the function of these candidate genes, however we conclude that our approach is effective at identifying novel players in haematopoiesis.

Type of Work:

Thesis (Masters by Research > M.Sc.)

Award Type:

Masters by Research > M.Sc.

Supervisor(s):