prikaz prve stranice dokumenta EFFECTS OF SIMULTANEOUS MANIPULATION OF DNA AND HISTONES ON THE EXPRESSION OF GENES INVOLVED IN EPITHELIAL-MESENCHYMAL TRANSITION USING CRISPR/dCas9 SYSTEM
Embargo period 2026-03-21
doctoral thesis
EFFECTS OF SIMULTANEOUS MANIPULATION OF DNA AND HISTONES ON THE EXPRESSION OF GENES INVOLVED IN EPITHELIAL-MESENCHYMAL TRANSITION USING CRISPR/dCas9 SYSTEM
Zagreb: University of Zagreb, Faculty of Science, 2024. urn:nbn:hr:217:444849
Foglar, Nika
University of Zagreb
Faculty of Science
Department of Biology

Institutional repository: Repository of the Faculty of Science

Cite this document

Foglar, N. (2024). EFFECTS OF SIMULTANEOUS MANIPULATION OF DNA AND HISTONES ON THE EXPRESSION OF GENES INVOLVED IN EPITHELIAL-MESENCHYMAL TRANSITION USING CRISPR/dCas9 SYSTEM (Doctoral thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:444849

Foglar, Nika. "EFFECTS OF SIMULTANEOUS MANIPULATION OF DNA AND HISTONES ON THE EXPRESSION OF GENES INVOLVED IN EPITHELIAL-MESENCHYMAL TRANSITION USING CRISPR/dCas9 SYSTEM." Doctoral thesis, University of Zagreb, Faculty of Science, 2024. https://urn.nsk.hr/urn:nbn:hr:217:444849

Foglar, Nika. "EFFECTS OF SIMULTANEOUS MANIPULATION OF DNA AND HISTONES ON THE EXPRESSION OF GENES INVOLVED IN EPITHELIAL-MESENCHYMAL TRANSITION USING CRISPR/dCas9 SYSTEM." Doctoral thesis, University of Zagreb, Faculty of Science, 2024. https://urn.nsk.hr/urn:nbn:hr:217:444849

Foglar, N. (2024). 'EFFECTS OF SIMULTANEOUS MANIPULATION OF DNA AND HISTONES ON THE EXPRESSION OF GENES INVOLVED IN EPITHELIAL-MESENCHYMAL TRANSITION USING CRISPR/dCas9 SYSTEM', Doctoral thesis, University of Zagreb, Faculty of Science, accessed 04 November 2024, https://urn.nsk.hr/urn:nbn:hr:217:444849

Foglar N. EFFECTS OF SIMULTANEOUS MANIPULATION OF DNA AND HISTONES ON THE EXPRESSION OF GENES INVOLVED IN EPITHELIAL-MESENCHYMAL TRANSITION USING CRISPR/dCas9 SYSTEM [Doctoral thesis]. Zagreb: University of Zagreb, Faculty of Science; 2024 [cited 2024 November 04] Available at: https://urn.nsk.hr/urn:nbn:hr:217:444849

N. Foglar, "EFFECTS OF SIMULTANEOUS MANIPULATION OF DNA AND HISTONES ON THE EXPRESSION OF GENES INVOLVED IN EPITHELIAL-MESENCHYMAL TRANSITION USING CRISPR/dCas9 SYSTEM", Doctoral thesis, University of Zagreb, Faculty of Science, Zagreb, 2024. Available at: https://urn.nsk.hr/urn:nbn:hr:217:444849

Metadata
TitleEFFECTS OF SIMULTANEOUS MANIPULATION OF DNA AND HISTONES ON THE EXPRESSION OF GENES INVOLVED IN EPITHELIAL-MESENCHYMAL TRANSITION USING CRISPR/dCas9 SYSTEM
Title (croatian)UTJECAJ ISTOVREMENE MANIPULACIJE DNA I HISTONA NA EKSPRESIJU GENA UKLJUČENIH U EPITELNO-MEZENHIMSKU TRANZICIJU PRIMJENOM SUSTAVA CRISPR/dCas9
AuthorNika Foglar
MentorVlatka Zoldoš (mentor)
Committee memberNataša Bauer (predsjednik povjerenstva)
Committee memberRosa Karlić (član povjerenstva)
Committee memberOliver Vugrek (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Biology)
Zagreb
Defense date and country2024-03-21, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Biology
Universal decimal classification
(UDC)		577 - Biochemistry. Molecular biology. Biophysics
Abstract
Interplay between epigenetic modifications is essential for regulation of gene expression. The adaptation of CRISPR/Cas9 technology for epigenetic research by introducing dead Cas9 (dCas9) fused to epigenetic effector domains (ED) has enabled significant advancements in this field. I expanded the CRISPR/dCas9 modular system with five histone modifying domains (G9a, G9a-me3, LSD1, KDM5A, macro1.1) and DNMT3L domain, as well as the multi-guide module to accommodate up to 11 gRNAs. From a set of candidate genes involved in epithelial to mesenchymal transition (EMT), ZEB1 and SNAI1 were selected for further epigenetic manipulations in HepG2 cell line. Their CpG islands were simultaneously targeted with dCas- DNMT3A combined with specific dCas9-ED fusions to understand the interplay of DNA methylation and histone modifications. The combinations had synergistic effect on gene expression and affected the EMT phenotype. Interplay between introduced modifications was also observed as a locus specific synergistic increase in DNA methylation, and was not disrupted by global histone acetylation increase after sodium butyrate treatment. The combinatorial epigenetic effects were not replicated in A549 cell line, suggesting specificity of epigenetic regulatory pathways.
Abstract (croatian)
Međudjelovanje epigenetičkih modifikacija je neophodno regulaciju ekspresije gena. Korištenje CRISPR/Cas9 tehnologije u epigenetičkim istraživanjima pomoću inaktivnog Cas9 (dCas9) fuzioniranog s epigenetičkim efektorskim domenama (ED) omogućilo je značajan napredak u ovom području. Proširila sam modularni sustav CRISPR/dCas9 s pet domena za modifikaciju histona (G9a, G9a-me3, LSD1, KDM5A, makro1.1) i domenom DNMT3L, te „multi-guide“ modul koji može primiti do 11 gRNA. Od nekoliko kandidat gena koji sudjeluju u epitelno-mezenhimskoj tranziciji (EMT), odabrala sam ZEB1 i SNAI1 za daljnje epigenetičke manipulacije u staničnoj liniji HepG2. Njihovi CpG otoci su istovremeno ciljani s DNMT3AdCas9 u kombinaciji s određenom dCas9-ED fuzijom kako bi se istražilo među-djelovanje DNA metilacije i modifikacija histona. Kombinacije su imale sinergistički učinak na ekspresiju gena i utjecale su na EMT fenotip. Simultano uvedene modifikacije pokazale su sinergistički efekt na razini metilacije DNA na određenom lokusu, koji nije bio osjetljiv na povećanje globalne acetilacije histona. Kombinacije dCas9 fuzija nisu djelovale na ekspresiju ciljanih gena u A549 staničnoj liniji, što ukazuje na specifičnost epigenetičkih regulatornih putova.
Keywords
Keywords (croatian)
Languageenglish
URN:NBNurn:nbn:hr:217:444849
Promotion2024
ProjectNumber: KK.01.1.1.04.0085
Title: Genomsko inženjerstvo i genska regulacija u staničnim linijama i modelnim organizacijama tehnologija CRISPR / Cas9 CasMouse
Title: Genomic engineering and gene regulation in the cell lines model organisms CRISPR / Cas9 CasMouse technology
Acronym: CRISPR/Cas9-CasMouse
Leader: Vlatka Zoldoš
Jurisdiction: Croatia
Funder: EK
Funding stream: EFRR
Study programmeTitle: Doctoral study
Study programme type: university
Study level: postgraduate
Academic / professional title: doktor/doktorica znanosti u području prirodnih znanosti (doktor/doktorica znanosti u području prirodnih znanosti)
Type of resourceText
File originBorn digital
Access conditionsEmbargoed access
Embargo expiration date: 2026-03-21
Terms of useLicence In copyright icon
Created on2024-04-02 10:13:19