prikaz prve stranice dokumenta Development of Dual Reporter Luciferase system for in vivo detection of miRNA target gene downregulation
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master's thesis
Development of Dual Reporter Luciferase system for in vivo detection of miRNA target gene downregulation
2016. urn:nbn:hr:193:078017
Žuža, Kristina
University of Rijeka
Faculty of Biotechnology and Drug Development

Cite this document

Žuža, K. (2016). Development of Dual Reporter Luciferase system for in vivo detection of miRNA target gene downregulation (Master's thesis). Rijeka: University of Rijeka. Retrieved from https://urn.nsk.hr/urn:nbn:hr:193:078017

Žuža, Kristina. "Development of Dual Reporter Luciferase system for in vivo detection of miRNA target gene downregulation." Master's thesis, University of Rijeka, 2016. https://urn.nsk.hr/urn:nbn:hr:193:078017

Žuža, Kristina. "Development of Dual Reporter Luciferase system for in vivo detection of miRNA target gene downregulation." Master's thesis, University of Rijeka, 2016. https://urn.nsk.hr/urn:nbn:hr:193:078017

Žuža, K. (2016). 'Development of Dual Reporter Luciferase system for in vivo detection of miRNA target gene downregulation', Master's thesis, University of Rijeka, accessed 19 November 2024, https://urn.nsk.hr/urn:nbn:hr:193:078017

Žuža K. Development of Dual Reporter Luciferase system for in vivo detection of miRNA target gene downregulation [Master's thesis]. Rijeka: University of Rijeka; 2016 [cited 2024 November 19] Available at: https://urn.nsk.hr/urn:nbn:hr:193:078017

K. Žuža, "Development of Dual Reporter Luciferase system for in vivo detection of miRNA target gene downregulation", Master's thesis, University of Rijeka, Rijeka, 2016. Available at: https://urn.nsk.hr/urn:nbn:hr:193:078017

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Metadata
TitleDevelopment of Dual Reporter Luciferase system for in vivo detection of miRNA target gene downregulation
Title (croatian)Razvoj dvostrukog luciferaznog reporter eseja za in vivo detekciju miRNA genskog utišavanja
AuthorKristina Žuža
MentorSerena Zacchigna (mentor)
MentorAntonija Jurak Begonja (komentor)
Committee memberAntonija Jurak Begonja (predsjednik povjerenstva)
Committee memberElitza Petkova Markova - Car (član povjerenstva)
Committee memberSandra Kraljević Pavelić (član povjerenstva)
GranterUniversity of Rijeka
(Faculty of Biotechnology and Drug Development)
Rijeka
Defense date and country2016-09-26, Croatia
Scientific / art field,
discipline and subdiscipline		BIOTECHNICAL SCIENCES
Biotechnology
Abstract
Most frequent global death causes are cardiovascular disorders such as cerebrovascular diseases, high blood pressure, congenital heart disease and coronary artery disease. Cardiovascular diseases often result in development of heart failure syndrome which in most cases leads to death within 5 years after diagnosis. Similar to heart failure, acute myocardial infarction is a silent murderer of today’s developed society due to sedentary life style and unhealthy habits such as smoking and unhealthy diet. Both disorders are characterized by heart cell remodeling and depletion in number of viable cardiomyocytes what is considered to be main obstacle in finding efficient therapy. Heart is an organ unable to respond to damage by self-renewal and therapy provided up to date is insufficient to increase number of viable CMs. Although many pharmaceuticals and surgical interventions are developed, all of them temporarily mend the heart. For this reason development of novel therapeutics is required among which miRNAs show to be very promising in boosting heart proliferation rate. MiRNA-199a-3p and 590-3p showed to significantly increase cardiomyocyte proliferation in vitro and in vivo and to preserve cardiac function after injury cause by mimicking myocardial infarction. Homer1 and CLIC5 genes are showed to be most down regulated by miRNA-199a-3p and 590-3p. To measure level of this downregulation more precisely and to show that this targeting is limited specifically to cardiomyocytes, not to other heart cells (cardiac fibroblasts, endothelial cells…) we decided to develop dual luciferase reporter model of monitoring miRNA targeting in vivo. System would take advantage of adeno-associated virus (AAV) vector tissue tropism, specifically transducing cardiomyocytes. Vector would carry transgene consisted of a 3’UTRs of Homer1 or CLIC5 gene together with luciferase (Renilla or firefly) sequence. Gene downregulation would be assesed by luciferase bioluminescence decrease after miRNA mimic intracardiac injection.
Abstract (croatian)
Kardiovaskularne bolesti kao što su visoki krvni tlak, cerebrovaskularna te koronarna oboljenja predstavljaju jedan od najčešćih uzroka smrti na globalnoj razini. Takva oboljenja često rezultiraju sindromom zatajenja srca koji najčešće dovodi do smrti pet godina nakon uspostavljanja dijagnoze. Slično kao i zatajenje srca, infarkt mokarda je tihi ubojica današnjeg razvijenog društva te je posljedica sjedilačkog načina života i nezdravih navika. Oba poremećaja su karaterizirana remodeliranjem srčanih stanica te smanjenim brojem funkcionalnih kardiomiocita što se smatra glavnom preprekom u pronalasku učinkovite terapije. Srce je organ kojeg karakterizira nemogućnost samo-obnove uslijed ozljede te su terapije razvijene do danas neučinkovite u povećanju broja funkcionalnih kardiomiocita. Razvijeni su mnogi korisni lijekovi i kirurški zahvati, ali oni često samo privremeno zaliječe srce. Sukladno tomu, nužan je razvoj novih terapija u liječenju srčanih oboljenja među kojima su se mikro RNA molekule pokazale kao vrlo obećavajuće u poticanju proliferacije kardiomiocita. Jedan od primjera su miRNA-199a-3p i miRNA-590-3p koje su pokazale da značajno povisuju razinu proliferacije kardiomiocita kako in vitro tako i in vivo. Također, pokazale su bitnu ulogu u očuvanju srčane funkcije nakon oštećenja izazvanog srčanim udarom. Dva gena koji su bili utišani u najvećoj mjeri od strane miRNA-199a-3p i miRNA-590-3p su Homer1 i CLIC5. Da bi u buduće što preciznije mjerili to utišavanje te kako bi dokazali da je ono ograničeno samo na kardiomiocite, a ne ostale srčane stanice (fibroblaste i endotelne stanice), odlučili smo razviti dvostruki luciferazni esej za in vivo detekciju miRNA genskog utišavanja. Sistem bi koristio prednost tkivne specifičnosti adeno-združenog viralnog vektora transducirajući isključivo kardiomiocite u srčanom tkivu. Vektor bi nosio transgen koji se sastoji od 3'UTR regije Homer1 ili CLIC5 gena zajedno sa sekvencom gena za luciferazu (Renilla ili firefly). Utišavanje gena bi se mjerilo smanjenjem luciferazne bioluminiscencije nakon intrakardijalne injekcije miRNA-199a-3p i miRNA-590-3p.
Keywords
Keywords (croatian)
Languageenglish
URN:NBNurn:nbn:hr:193:078017
Study programmeTitle: Biotechnology in medicine
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra biotehnologije u medicini (magistar/magistra biotehnologije u medicini)
Type of resourceText
File originBorn digital
Access conditionsClosed access
Terms of useLicence In copyright icon
Created on2016-11-02 10:48:53