Potential downstream targets of the polyamine pathway and their roles in embryonic stem cell renewal

• Main Author: Fong, Koon Kit
• Other Authors: Leah Vardy
• Format: Final Year Project
• Language: English
• Published: 2015
• Subjects: DRNTU::Science::Biological sciences::Molecular biology
• Online Access: http://hdl.handle.net/10356/63170
• Institution: Nanyang Technological University

Embryonic stem cells hold great potential for regenerative medicine due to their ability to self-renew and differentiate into cells of all embryonic cell lineages. Polyamines are cationic molecules consisting of putrescine, spermidine and spermine A polyamine synthesis regulator, adenosylmethionine decarboxylase 1 (AMD1) was shown to be essential for embryonic stem cell self-renewal. A microarray was performed to identify genes that were downregulated on ESC differentiation while upregulated upon AMD1 overexpression. These genes are potential downstream targets of the polyamine pathway that could regulate embryonic stem cell self-renewal. Functional assays of overexpression and knockdown were performed for the candidate genes of Aqp3 and Zmat4. Aqp3 and Zmat4 isoforms were overexpressed and knocked down using siRNA. Core pluripotency genes and differentiation markers were examined by qRT-PCR while pluripotency was tested with alkaline phosphatase staining. Overexpression and knockdown results suggest that Aqp3 and Zmat4 did not have a significant impact on pluripotency. However, more experiments are required to have a definitive conclusion.