Korištenje CRISPR/Cas9 metode za inaktivaciju tirozil-DNA fosfodiesteraze 1 (TDP1) u zebrici (Danio rerio)

undergraduate thesis

• Title (english): Using CRISPR/Cas9 method for Tyrosyl-DNA phosphodiesterase 1 inactivation in zebrafish (Danio rerio)
• Author: Arijana Dubović
• Mentor: Igor Stuparević (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Chemistry and Biochemistry); (Laboratory for Biochemistry); Zagreb
• Defense date and country: 2020-07-10, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Tirozil-DNA fosfodiesteraza 1 (Tdp1) enzim je čija je in vivo uloga slabo istražena. Do sada je potvrđeno da Tdp1 in vivo sudjeluje u popravku kovalentno vezanih DNA-topoizomeraza 1 (Top1cc) oštećenja. Međutim, potrebno je prikupiti više informacija o regulaciji, međudjelovanju s drugim putevima popravka te postojanju dodatnih supstrata za Tdp1. Top1cc oštećenje za stanicu je često letalno i danas se intenzivno istražuju citostatici koji, inducirajući nakupljanje Top1cc sprječavaju širenje tumorskih stanica. Za uspješan razvoj ovih lijekova bitno je razumjeti način djelovanja Tdp1 enzima kako bi se mogao minimizirati efekt popravka induciranih oštećenja. Razumijevanje funkcije Tdp1 enzima bio bi veliki korak i za liječenje SCAN1 poremećaja uzrokovanog mutacijama u genu za Tdp1. Cilj ovog rada omogućiti je daljnja istraživanja Tdp1 enzima in vivo u čiju je svrhu sintetizirana RNA vodilja koja će se koristiti za inaktivaciju Tdp1 CRISPR/Cas9 metodom u zebrici (Danio rerio).

Abstract (english)

Tyrosyl-DNA phosphodiesterase 1 (Tdp1) is an enzyme whose function in vivo is poorly understood. It's proven that Tdp1 plays a role in DNA-topoisomerase1 (Top1cc) crosslink repair in vivo. Yet more information is needed on regulation, interplay with other repair pathways and identification of additional substrate. Top1cc occurance is often deadly for cells, and intenstive research is done nowadays to develop citostatic medications which induce this type of damage to fight cancer cells. For a successful drug development, Tdp1 function should be well understood to minimise the repair of deliberately induced Top1ccs. In addition, understanding the Tdp1 function would make a big step forward in treating SCAN1 disorder which is caused by mutations in tdp1 gene. This beachlor thesis has a purpose to enable further research on Tdp1 by creating a single guide RNA which will be used for a tdp1 gene knock-out using CRISPR/Cas9 method in zebrafish (Danio rerio).

• Keywords: CRISPR/Cas9; Danio rerio; DPC; Top1cc; Tdp1
• Keywords (english): CRISPR/Cas9; Danio rerio; DPC; Tdp1; TOP1cc
• Language: croatian
• URN:NBN: urn:nbn:hr:159:680098
• Study programme: Title: Biotechnology; Study programme type: university; Study level: undergraduate; Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2020-11-10 11:32:08