MBD1 mediated enrichment of methylated DNA from cell lines deficient in S-adenosylhomocysteine hydrolase (AHCY)

Polović, Mirjana (2013) MBD1 mediated enrichment of methylated DNA from cell lines deficient in S-adenosylhomocysteine hydrolase (AHCY). Diploma thesis, Faculty of Science > Department of Biology.

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Abstract

Methylation plays an important role in regulating biological processes. Abnormalities in DNA methylation are involved in the etiology of various human disorders. S-adenosylhomocysteine hydrolase (AHCY) deficiency is a novel human disease, discovered in Croatian population in 2004. The disease results in striking elevations of S-adenosylhomocysteine (adoHcy) and S-adenosylmethionine (adoMet) in blood and various tissues. AHCY efficiently removes adoHcy, the by-product of transmethylation reactions and potent DNA methyltransferase inhibitor. Thus, AHCY plays a critical role in regulation of biological methylation. The goal of my research was to evaluate AHCY deficiency derived changes in DNA methylation. To do so, DNA was isolated from human liver tumor cell line, HepG2, which has been silenced for AHCY expression using RNA interference technology. DNA methylation status was studied on certain imprinted loci and confirmed by bisulfite sequencing. Loci with significant changes in methylation status may serve as biomarkers for disease detection.

Item Type: Thesis (Diploma thesis)
Keywords: AHCY deficiency, bisulfite sequencing, DNA methylation, HepG2 cell line, imprinted loci
Supervisor: Vugrek, Oliver
Co-supervisor: Zoldoš, Vlatka
Date: 2013
Number of Pages: 59
Subjects: NATURAL SCIENCES > Biology
Divisions: Faculty of Science > Department of Biology
Depositing User: Grozdana Sirotic
Date Deposited: 12 Feb 2014 13:11
Last Modified: 22 Sep 2014 19:17
URI: http://digre.pmf.unizg.hr/id/eprint/420

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