prikaz prve stranice dokumenta Unos i učinci genskih preinaka posredovanih lentivirusnim vektorom u mišjem mozgu
Embargo period 2025-04-17
doctoral thesis
Unos i učinci genskih preinaka posredovanih lentivirusnim vektorom u mišjem mozgu
Zagreb: University of Zagreb, School of Medicine, 2024. urn:nbn:hr:105:277371
Skukan, Laura
University of Zagreb
School of Medicine

Institutional repository: Dr Med - University of Zagreb School of Medicine Digital Repository

Cite this document

Skukan, L. (2024). Unos i učinci genskih preinaka posredovanih lentivirusnim vektorom u mišjem mozgu (Doctoral thesis). Zagreb: University of Zagreb, School of Medicine. Retrieved from https://urn.nsk.hr/urn:nbn:hr:105:277371

Skukan, Laura. "Unos i učinci genskih preinaka posredovanih lentivirusnim vektorom u mišjem mozgu." Doctoral thesis, University of Zagreb, School of Medicine, 2024. https://urn.nsk.hr/urn:nbn:hr:105:277371

Skukan, Laura. "Unos i učinci genskih preinaka posredovanih lentivirusnim vektorom u mišjem mozgu." Doctoral thesis, University of Zagreb, School of Medicine, 2024. https://urn.nsk.hr/urn:nbn:hr:105:277371

Skukan, L. (2024). 'Unos i učinci genskih preinaka posredovanih lentivirusnim vektorom u mišjem mozgu', Doctoral thesis, University of Zagreb, School of Medicine, accessed 22 November 2024, https://urn.nsk.hr/urn:nbn:hr:105:277371

Skukan L. Unos i učinci genskih preinaka posredovanih lentivirusnim vektorom u mišjem mozgu [Doctoral thesis]. Zagreb: University of Zagreb, School of Medicine; 2024 [cited 2024 November 22] Available at: https://urn.nsk.hr/urn:nbn:hr:105:277371

L. Skukan, "Unos i učinci genskih preinaka posredovanih lentivirusnim vektorom u mišjem mozgu", Doctoral thesis, University of Zagreb, School of Medicine, Zagreb, 2024. Available at: https://urn.nsk.hr/urn:nbn:hr:105:277371

Metadata
TitleUnos i učinci genskih preinaka posredovanih lentivirusnim vektorom u mišjem mozgu
Title (english)Application and effects of gene modifications mediated by a lentviral vector in the mouse brain
AuthorLaura Skukan
MentorSrećko Gajović (mentor)
Committee memberŽeljka Krsnik (predsjednik povjerenstva)
Committee memberDubravka Švob Štrac (član povjerenstva)
Committee memberNataša Jovanov Milošević (član povjerenstva)
Committee memberDinko Mitrečić (član povjerenstva)
Committee memberGoran Sedmak (član povjerenstva)
GranterUniversity of Zagreb
School of Medicine
Zagreb
Defense date and country2024-04-16, Croatia
Scientific / art field,
discipline and subdiscipline		BIOMEDICINE AND HEALTHCARE
Basic Medical Sciences
Neuroscience
Universal decimal classification
(UDC)		61 - Medical sciences
Abstract
Unos genetskog materijala i genskih preinaka važan je pristup za proučavanje i liječenje genetskih i drugih bolesti. Lentivirusni vektori ističu se kao dominatan sustav dostave u ciljane stanične linije središnjeg živčanog sustava. Glavni cilj ovog istraživanja bio je utvrditi transdukciju neurona i astrocita za njih specifično dizajniranim lentivirusnim vektorom, procjeniti uzrokuje li dostavljen sustav shRNA utišavanje gena Spry2 te utvrditi učinak utišavanja gena Spry2 nakon izazivanja ishemijskog moždanog udara u mišjem modelu. Potvrđena je uspješna transdukcija stanica lentivirusnim vektorom te je analizom kolokalizacije pokazano da lentivirusni vektori s ovojnicom VSV-G transduciraju neurone dok Tet regulirani lentivirusni vektori sa specifičnim promotorom za astrocite i ovojnicom Mokola transduciraju astrocite. Procjenom aktivacije mikroglije i receptora TLR2 pokazano je da unos lentivirusnog vektora uzrokuje kratkotrajnu aktivaciju upalnog odgovora koja je ograničena na područje unosa. Analizom genske ekspresije potvrđeno je utišavanje gena Spry2 u injektiranim regijama mišjeg mozga. Međutim, njegova smanjena ekspresija nije pridonijela značajnom smanjenju volumena ishemijske lezije i funkcionalnom oporavku nakon ishemijskog moždanog udara. Ovo istraživanje potvrdilo je da su primjenjene strategije dizajna lentivirusnih vektora učinkovit sustav dostave koji omogućava ograničenu transdukciju stanica od interesa i uspješno utišavanje gena Spry2.
Abstract (english)
The delivery of genetic material or gene modification is an important approach for the study and treatment of genetic and other diseases. Lentiviral vectors are a dominant delivery system to target cell lines of the central nervous system. The aim of this study was to determine the transduction of target cells with a specifically designed lentiviral vector for the neuron and astrocyte transduction, to assess whether the delivered shRNA system causes silencing of Spry2, and to examine the effect of Spry2 silencing after ischemic stroke induction in a mouse model. Successful cell transduction with the lentiviral vector was confirmed, and colocalization analysis confirmed that VSV-G pseudotyped lentiviral vectors transduce neurons, while Tet-regulated Mokola pseudotyped lentiviral vector with astrocyte-specific promoter transduce astrocytes. Furthermore, lentiviral vector causes a short-term activation of the inflammatory response, which was restricted near the area of injection site. Silencing of the Spry2 was confirmed in the injected regions of the mouse brain. However, downregulation of Spry2 did not reduced the volume of ischemic lesion and contribute to the functional recovery after ischemia. This study confirmed that the applied design strategies of lentiviral vector are an efficient delivery system that allows restricted transduction of cells of interest and successful Spry2 silencing.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:105:277371
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: Srećko Gajović
Jurisdiction: Croatia
Funder: EK
Funding stream: EFRR
Study programmeTitle: Neuroscience
Study programme type: university
Study level: postgraduate
Academic / professional title: doktor/doktorica znanosti, područje biomedicine i zdravstvo (doktor/doktorica znanosti, područje biomedicine i zdravstvo)
Type of resourceText
Extent120 str.
File originBorn digital
Access conditionsEmbargoed access
Embargo expiration date: 2025-04-17
Terms of useLicence In copyright icon
Created on2024-04-19 10:57:13