Obogaćivanje posredovano MBD1 metilirane DNA iz staničnih linija s nedostatnom aktivnosti S-adenozilhomocistein hidrolaze (AHCY)

master's thesis

• Title (english): MBD1 mediated enrichment of methylated DNA from cell lines deficient in S-adenosylhomocysteine hydrolase (AHCY)
• Author: Mirjana Polović
• Mentor: Oliver Vugrek (mentor)
• Mentor: Vlatka Zoldoš (mentor)
• Committee member: Vlatka Zoldoš (predsjednik povjerenstva)
• Committee member: Oliver Vugrek (član povjerenstva)
• Committee member: Ana Galov (član povjerenstva)
• Committee member: Višnja Besendorfer (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2013-02-12, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Metilacija igra važnu ulogu u regulaciji bioloških procesa. Nepravilnosti u DNA metilaciji su uključene u nastanak različitih poremećaja zdravlja u ljudi. Nedostatnost S-adenozil homocistein hidrolaze (AHCY) je nova bolest, otkrivena u Hrvatskoj populaciji 2004. godine. Bolest rezultira u upečatljivom povišenju S-adenozil homocisteina (adoHcy) i S-adenozil metionina (adoMet) u krvi i različitim tkivima. AHCY uspješno uklanja adoHcy, nusprodukt transmetilacijskih reakcija i moćan inhibitor DNA metiltransferaza. Dakle, AHCY igra ključnu ulogu u regulaciji biološke metilacije. Cilj mog istraživanja bio je procijeniti promijene u DNA metilaciji nastale zbog nedostatnosti AHCY. Zato je DNA izolirana iz stanične linije ljudskog tumora jetre, HepG2, koja je utišana u AHCY ekspresiji korištenjem RNA interferencijske tehnologije. DNA metilacijski status je proučen na odabranim utisnutim lokusima te utvrđen bisulfitnim sekvenciranjem. Lokusi sa značajnim promjenama u metilacijskom statusu se mogu koristiti kao biomarkeri za detekciju bolesti.

Abstract (english)

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.

• Keywords: nedostatnost AHCY; bisulfitno sekvenciranje; DNA metilacija; HepG2 stanična linija; utisnuti lokusi
• Keywords (english): AHCY deficiency; bisulfite sequencing; DNA methylation; HepG2 cell line; imprinted loci
• Language: croatian
• URN:NBN: urn:nbn:hr:217:539752
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-04-27 12:33:14