prikaz prve stranice dokumenta Ciljano uklanjanje histonske oznake H3K4me3 u promotoru gena B4GALT1 upotrebom fuzijskog proteina RIOX1-dSpCas9
Embargo period 2026-03-18
master's thesis
Ciljano uklanjanje histonske oznake H3K4me3 u promotoru gena B4GALT1 upotrebom fuzijskog proteina RIOX1-dSpCas9
Zagreb: University of Zagreb, Faculty of Science, 2024. urn:nbn:hr:217:570262
Zandona, Gea
University of Zagreb
Faculty of Science
Department of Biology

Institutional repository: Repository of the Faculty of Science

Cite this document

Zandona, G. (2024). Ciljano uklanjanje histonske oznake H3K4me3 u promotoru gena B4GALT1 upotrebom fuzijskog proteina RIOX1-dSpCas9 (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:570262

Zandona, Gea. "Ciljano uklanjanje histonske oznake H3K4me3 u promotoru gena B4GALT1 upotrebom fuzijskog proteina RIOX1-dSpCas9." Master's thesis, University of Zagreb, Faculty of Science, 2024. https://urn.nsk.hr/urn:nbn:hr:217:570262

Zandona, Gea. "Ciljano uklanjanje histonske oznake H3K4me3 u promotoru gena B4GALT1 upotrebom fuzijskog proteina RIOX1-dSpCas9." Master's thesis, University of Zagreb, Faculty of Science, 2024. https://urn.nsk.hr/urn:nbn:hr:217:570262

Zandona, G. (2024). 'Ciljano uklanjanje histonske oznake H3K4me3 u promotoru gena B4GALT1 upotrebom fuzijskog proteina RIOX1-dSpCas9', Master's thesis, University of Zagreb, Faculty of Science, accessed 26 November 2024, https://urn.nsk.hr/urn:nbn:hr:217:570262

Zandona G. Ciljano uklanjanje histonske oznake H3K4me3 u promotoru gena B4GALT1 upotrebom fuzijskog proteina RIOX1-dSpCas9 [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2024 [cited 2024 November 26] Available at: https://urn.nsk.hr/urn:nbn:hr:217:570262

G. Zandona, "Ciljano uklanjanje histonske oznake H3K4me3 u promotoru gena B4GALT1 upotrebom fuzijskog proteina RIOX1-dSpCas9", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2024. Available at: https://urn.nsk.hr/urn:nbn:hr:217:570262

Metadata
TitleCiljano uklanjanje histonske oznake H3K4me3 u promotoru gena B4GALT1 upotrebom fuzijskog proteina RIOX1-dSpCas9
Title (english)Targeting the fusion protein RIOX1-dSpCas9 to the promoter region of B4GALT1 for removal of the histone mark H3K4me3
AuthorGea Zandona
MentorVlatka Zoldoš (mentor)
Committee memberVlatka Zoldoš (predsjednik povjerenstva)
Committee memberMirjana Pavlica (član povjerenstva)
Committee memberMaja Matulić (član povjerenstva)
Committee memberPetra Korać (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Biology)
Zagreb
Defense date and country2024-03-18, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Biology
Abstract
Šest epigenetičkih mehanizama simultanim djelovanjem regulira gensku ekspresiju, a njihove su interakcije, zastupljenosti i efekti na transkripcijski status određenog gena specifični u odnosu na kromatinski „okoliš“. Cilj istraživanja bio je ukloniti histonsku oznaku H3K4me3 u promotorskoj regiji gena B4GALT1 pomoću fuzijskog proteina RIOX1-dSpCas9 navođenog na specifičnu regiju sa šest različitih molekula sgRNA (od engl. single guide RNA). Također, cilj je bio dizajnirati testove za analizu stupnja metilacije pirosekvenciranjem ove regije. Metodom kromatinske precipitacije CUT&RUN, nakon koje slijedi qPCR, utvrđeno je da nije došlo do statistički značajnog smanjenja u količini histonske oznake H3K4me3 kao ni do povećanja stupnja metilacije DNA u analiziranoj regiji. Metodom kvantitativnog PCR je utvrđeno da nije došlo do promjene u genskoj ekspresiji. Gen B4GALT1 ima složenu genetičku i epigenetičku regulaciju, a efekt fuzijskog proteina RIOX1-dSpCas9 mogao je ostati nezapažen zbog uskog područja koje je analizirano, ali i zbog već prisutnih epigenetičkih modifikacija i strukture kromatina koji su mogli onemogućiti njegovo vezanje ili katalitičku aktivnost.
Abstract (english)
Gene expression is regulated by six epigenetic mechanisms that work simultaneously. Their interactions as well as their effects on the transcriptional status of a certain gene are specific and depend on chromatin context. The goal of this research was to remove the histone mark H3K4me3 from the promoter region of the B4GALT1 gene using the fusion protein RIOX1-dSpCas9, which was guided to a specific region by six different sgRNA molecules. Also, the goal was to design assays for pyrosequencing in order to analyze DNA methylation level in this region. The CUT&RUN chromatin precipitation method followed with qPCR revealed no statistically significant change in the amount of histone mark H3K4me3 nor in DNA methylation level of the analyzed region. Quantitative PCR showed no change in the the B4GALT1 gene transcriptional activity. The B4GALT1 gene has complex genetic and epigenetic regulation, and the effect of the RIOX1-dSpCas9 fusion protein could have gone unnoticed due to the narrow region in the gene promoter that was analyzed. Also, the already present epigenetic modifications and chromatin structure in the targeted region could have prevent the binding or catalytic activity of RIOX1-dSpCas9.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:217:570262
Study programmeTitle: Molecular Biology-University graduate programme
Study programme type: university
Study level: graduate
Academic / professional title: sveučilišni/a magistar/magistra molekularne biologije (sveučilišni/a magistar/magistra molekularne biologije)
Type of resourceText
File originBorn digital
Access conditionsEmbargoed access
Embargo expiration date: 2026-03-18
Terms of useLicence In copyright icon
Created on2024-04-02 09:41:07