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master's thesis
Utjecaj narušavanja gena HBS1L, PELO, NONO, SND1 na mRNA koja kodira mutirani protein S-adenozilhomocistein-hidrolazu
Zagreb: University of Zagreb, Faculty of Science, 2020. urn:nbn:hr:217:244028
Šimunić, Ena
University of Zagreb
Faculty of Science
Department of Chemistry

Institutional repository: Repository of the Faculty of Science

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Šimunić, E. (2020). Utjecaj narušavanja gena HBS1L, PELO, NONO, SND1 na mRNA koja kodira mutirani protein S-adenozilhomocistein-hidrolazu (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:244028

Šimunić, Ena. "Utjecaj narušavanja gena HBS1L, PELO, NONO, SND1 na mRNA koja kodira mutirani protein S-adenozilhomocistein-hidrolazu." Master's thesis, University of Zagreb, Faculty of Science, 2020. https://urn.nsk.hr/urn:nbn:hr:217:244028

Šimunić, Ena. "Utjecaj narušavanja gena HBS1L, PELO, NONO, SND1 na mRNA koja kodira mutirani protein S-adenozilhomocistein-hidrolazu." Master's thesis, University of Zagreb, Faculty of Science, 2020. https://urn.nsk.hr/urn:nbn:hr:217:244028

Šimunić, E. (2020). 'Utjecaj narušavanja gena HBS1L, PELO, NONO, SND1 na mRNA koja kodira mutirani protein S-adenozilhomocistein-hidrolazu', Master's thesis, University of Zagreb, Faculty of Science, accessed 22 November 2024, https://urn.nsk.hr/urn:nbn:hr:217:244028

Šimunić E. Utjecaj narušavanja gena HBS1L, PELO, NONO, SND1 na mRNA koja kodira mutirani protein S-adenozilhomocistein-hidrolazu [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2020 [cited 2024 November 22] Available at: https://urn.nsk.hr/urn:nbn:hr:217:244028

E. Šimunić, "Utjecaj narušavanja gena HBS1L, PELO, NONO, SND1 na mRNA koja kodira mutirani protein S-adenozilhomocistein-hidrolazu", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2020. Available at: https://urn.nsk.hr/urn:nbn:hr:217:244028

Metadata
TitleUtjecaj narušavanja gena HBS1L, PELO, NONO, SND1 na mRNA koja kodira mutirani protein S-adenozilhomocistein-hidrolazu
Title (english)The effect of HBS1L, PELO, NONO and SND1 knock-out on mutated S-adenosylhomocysteine hydrolase mRNA
AuthorEna Šimunić
MentorOliver Vugrek (mentor)
Committee memberJasmina Rokov Plavec (predsjednik povjerenstva)
Committee memberIvan Kodrin (član povjerenstva)
Committee memberSnežana Miljanić (član povjerenstva)
Committee memberIvica Đilović (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Chemistry)
Zagreb
Defense date and country2020-02-17, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Chemistry
Abstract
S-adenozilhomocistein-hidrolaza (SAHH) je enzim koji katalizira reakciju hidrolize Sadenozil- L-homocisteina na adenozin i homocistein. SAHH ima ključnu ulogu u održavanju metilacijskog potencijala u stanici te njegov nedostatak dovodi do teškog metaboličkog poremećaja. Preliminarni podaci pokazuju da jedna od dosad opisanih mutacija (c.982 T>G), uz smanjenje aktivnosti enzima uzrokuje i smanjenje razine SAHH mRNA uz smanjenu razinu proteina SAHH u stanicama pacijenta. Pretpostavlja se da je mehanizam razgradnje uslijed zastajanja ribosoma tijekom translacije vjerojatni mehanizam razgradnje te mRNA. U svrhu potvrde hipoteze generirane su četiri stanične linije u kojima su metodom CRISPRCas9 pojedinačno narušeni geni HBS1L, PELO, NONO i SND1, potencijalni ključni čimbenici u razgradnji mRNA koja kodira mutirani protein SAHH, te je analiziran utjecaj njihovog narušavanja na razinu ekspresije te mRNA. Rezultati analize razine SAHH mRNA u staničnim linijama s narušenim genima metodom RT-qPCR pokazuju da jedino u staničnoj liniji s narušenim genom NONO dolazi do povrata razine ekspresije mutiranog transkripta.
Abstract (english)
S-adenosylhomocystein hydrolase (AHCY) is an enzyme that catalyses hydrolysis of Sadenosyl- L-homocysteine to adenosine and homocysteine. AHCY has the key role in maintaining cells' methylation potential, and its deficiency leads to severe metabolic disorder. Preliminary data shows that one of the mutations described so far (c.982 T>G) leads to decrease of the enzyme function and also causes a decrease of AHCY mRNA levels together with decreased AHCY protein levels in patient cells. It can be assumed that no-go decay is the probable mechanism of AHCY mRNA decay. To test this hypothesis, four knock-out cell lines have been generated. Using a CRISPR-Cas9 mediated approach, four genes, e.g. HBS1L, PELO, NONO and SND1 have been proposed as potential key factors in AHCY mRNA decay. Subsequently, all aforementioned genes have been individually knocked-out, and knock-out effects on AHCY mRNA expression levels have been analysed. RT-qPCR results show an expected rise in AHCY mRNA levels only in NONO knock-out cell line.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:217:244028
Study programmeTitle: Graduate university studies in chemistry; specializations in: Research programme
Course: Research programme
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra kemije (magistar/magistra kemije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Embargo expiration date: 2020-08-17
Terms of useLicence In copyright icon
Created on2020-02-21 12:12:59