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master's thesis
Ciljana mutageneza gena SPRTN i Tdp2a u zebrici (Danio rerio) pomoću sustava CRISPR
Zagreb: University of Zagreb, Faculty of Science, 2021. urn:nbn:hr:217:045651
Eršetić, Matea
University of Zagreb
Faculty of Science
Department of Biology

Institutional repository: Repository of the Faculty of Science

Cite this document

Eršetić, M. (2021). Ciljana mutageneza gena SPRTN i Tdp2a u zebrici (Danio rerio) pomoću sustava CRISPR (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:045651

Eršetić, Matea. "Ciljana mutageneza gena SPRTN i Tdp2a u zebrici (Danio rerio) pomoću sustava CRISPR." Master's thesis, University of Zagreb, Faculty of Science, 2021. https://urn.nsk.hr/urn:nbn:hr:217:045651

Eršetić, Matea. "Ciljana mutageneza gena SPRTN i Tdp2a u zebrici (Danio rerio) pomoću sustava CRISPR." Master's thesis, University of Zagreb, Faculty of Science, 2021. https://urn.nsk.hr/urn:nbn:hr:217:045651

Eršetić, M. (2021). 'Ciljana mutageneza gena SPRTN i Tdp2a u zebrici (Danio rerio) pomoću sustava CRISPR', Master's thesis, University of Zagreb, Faculty of Science, accessed 22 November 2024, https://urn.nsk.hr/urn:nbn:hr:217:045651

Eršetić M. Ciljana mutageneza gena SPRTN i Tdp2a u zebrici (Danio rerio) pomoću sustava CRISPR [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2021 [cited 2024 November 22] Available at: https://urn.nsk.hr/urn:nbn:hr:217:045651

M. Eršetić, "Ciljana mutageneza gena SPRTN i Tdp2a u zebrici (Danio rerio) pomoću sustava CRISPR", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2021. Available at: https://urn.nsk.hr/urn:nbn:hr:217:045651

Metadata
TitleCiljana mutageneza gena SPRTN i Tdp2a u zebrici (Danio rerio) pomoću sustava CRISPR
Title (english)Targeted mutagenesis of SPRTN and Tdp2a genes in zebrafish (Danio rerio) using the CRISPR system
AuthorMatea Eršetić
MentorMarta Popović (mentor)
MentorIvana Ivančić Baće (sumentor)
Committee memberIvana Ivančić Baće (predsjednik povjerenstva)
Committee memberIvana Šola (član povjerenstva)
Committee memberDubravka Hranilović (član povjerenstva)
Committee memberTomislav Ivanković (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Biology)
Zagreb
Defense date and country2021-09-27, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Biology
Abstract
Unakrsna vezanja DNA-protein nastaju uspostavom ireverzibilnih kovalentnih veza između DNA i proteina, a njihovo je uklanjanje neophodno za očuvanje integriteta staničnog genoma. Metaloproteaza Sprtn sudjeluje u proteolitičkoj razgradnji adukata histona i topoizomeraza, a nastali peptidni ostatci topoizomeraze II supstrat su za tirozil-DNA fosfodiesterazu 2 (Tdp2). Mutacije navedenih proteina smanjuju sposobnost popravka oštećenja DNA, a poznata je mutacija Y117C koja uzrokuje sindrom Rujis-Aalfs u ljudi, kojeg karakteriziraju preuranjeno starenje i razvoj hepatocelularnog karcinoma u mladoj dobi. Uvođenjem mutacije L147A u katalitičku regiju gena sprtn ribe zebrice (Danio rerio) pomoću sustava CRISPR/Cas12a, izradila sam animalni model sa smanjenom efikasnošću popravka unakrsnih vezanja DNAprotein za proučavanje funkcije proteina Sprtn. Također, selektirala sam dva utemeljitelja s pomakom okvira čitanja u genu tdp2a zebrice nakon primjene sustava CRISPR/Cas9 i uzgojila njihovo potomstvo. Uzgoj homozigotnih linija za navedene mutacije i mogućnost stvaranja dvostrukih mutanata poboljšat će razumijevanje uloga proteina Tdp2a i Sprtn u popravku oštećenja DNA te imati potencijalne primjene u liječenju bolesti čovjeka uzrokovanih mutacijama u navedenim genima.
Abstract (english)
DNA-protein crosslinks occur by establishing irreversible covalent bonds between DNA and proteins, and their removal is necessary to preserve the integrity of the cellular genome. The metalloprotease Sprtn participates in the proteolytic degradation of histone and topoisomerase adducts, and the resulting peptide residues of topoisomerase II are substrates for tyrosyl-DNA phosphodiesterase 2 (Tdp2). Mutations in these proteins reduce the ability to repair DNA damage, and the Y117C mutation is known to cause Rujis-Aalfs syndrome in humans, which is characterized by premature aging and the development of hepatocellular carcinoma at a young age. By introducing the L147A mutation into the catalytic region of the zebrafish (Danio rerio) sprtn gene using the CRISPR/Cas12a system, I developed an animal model with reduced DNA-protein crosslink repair efficiency to study Sprtn protein function. Also, I selected two founders with a frameshift in the zebrafish tdp2a gene after applying the CRISPR/Cas9 system and bred their offspring. The cultivation of homozygous lines for these mutations and the ability to form double mutants will improve understanding of the roles of Tdp2a and Sprtn proteins in DNA damage repair and have potential applications in the treatment of human diseases caused by mutations in these genes.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:217:045651
Study programmeTitle: Molecular Biology
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Embargo expiration date: 2023-09-27
Terms of useLicence In copyright icon
Created on2022-01-25 12:33:35