20.500.12556/RUL-186543 Analiza regulacije izražanja gena Pla2g4e in njegovih kolokaliziranih paralogov pri miši (Mus musculus) Analysis of the regulation of Pla2g4e gene expression and colocalized paralogs in the mouse (Mus musculus) Debelost je kompleksna bolezen, pri kateri pomembno vlogo igrajo okoljski in genetski dejavniki. Za preučevanje te bolezni se pogosto uporabljajo mišji modeli, pri čemer selekcijski liniji debelih (FLI) in vitkih (FHI) miši predstavljata enega najboljših modelov poligenske debelosti. V genskem skupku kolokaliziranih citosolnih fosfolipaz A2 na kromosomu 2, ki vključuje gene Pla2g4b, Pla2g4d, Pla2g4e in Pla2g4f, je bil Pla2g4e predhodno identificiran kot kandidatni gen za debelost. Z bioinformatskimi analizami so bile identificirane regulatorne regije, ki so potencialno odgovorne za diferencialno izražanje tega gena med debelo in vitko linijo miši. Prvi cilj te magistrske naloge je bil preveriti, ali imajo variacije v zaporedju DNA v izbranih regijah vpliv na izražanje gena Pla2g4e, za kar smo uporabili luciferazni reporterski test. Rezultati za testirani regiji niso pokazali statistično značilnih razlik v regulatorni aktivnosti med debelo in vitko linijo, pri čemer trend v regiji F1/R1 nakazuje možen vpliv alelov. Drugi cilj je bil analizirati vzorce izražanja vseh štirih kolokaliziranih paralognih genov in ugotoviti, ali so podvrženi koekspresiji. Analiza qPCR v naboru tkiv miši linije C57BL/6, podlinije J je pokazala, da imajo kolokalizirani paralogni geni izrazito različne, tkivno specifične profile izražanja. Korelacijska analiza po Spearmanu je pokazala statistično značilno korelacijo med ekspresijskim profilom Pla2g4b in vsemi tremi ostalimi geni, medtem ko korelacije med preostalimi pari niso bile statistično značilne. Rezultati podpirajo funkcionalno divergenco kolokaliziranih paralogov ter nakazujejo, da diferencialno izražanje gena Pla2g4e ni zgolj posledica skupne regulacije celotnega genskega skupka, temveč odraža njegovo specifično regulacijo. Obesity is a complex disease subject to numerous environmental and genetic influences. Mouse models are frequently used to study this disease, with the fat (FLI) and lean (FHI) selection lines representing one of the best models of polygenic obesity. Within the gene cluster of co-localised cytosolic phospholipases A2 on chromosome 2, comprising the genes Pla2g4b, Pla2g4d, Pla2g4e and Pla2g4f, the gene Pla2g4e was previously identified as a candidate obesity gene. Bioinformatic analyses identified regulatory regions potentially responsible for the differential expression of this gene between the fat and lean mouse lines. The first aim of this master’s thesis was to determine whether DNA sequence variations in selected regions influence the expression of the Pla2g4e gene, for which a luciferase reporter assay was used. Results for the two tested regions did not show statistically significant differences in regulatory activity between the Fat and Lean lines, although a trend in the F1/R1 region suggests a possible allelic effect. The second objective was to analyse the expression patterns of all four co-localised paralogous genes and determine whether they are subject to co-expression. qPCR analysis across a panel of tissues from the J substrain of C57BL/6 mouse line revealed that the colocalized paralogous genes display markedly distinct tissue-specific expression profiles. Spearman correlation analysis showed a statistically significant correlation between the expression profile of Pla2g4b and all three other genes, while correlations between the other gene pairs were not statistically significant. The results support functional divergence of the colocalized paralogs and suggest that the differential expression of Pla2g4e is not merely a consequence of shared regulation of the entire gene cluster but reflects its specific regulatory control. genetika Pla2g4b Pla2g4d Pla2g4e Pla2g4f izražanje genov paralogni geni koekspresija debelost laboratorijske miši vitka linija debela linija genetics Pla2g4b Pla2g4d Pla2g4e Pla2g4f gene expression paralogous genes co-expression obesity laboratory mice lean line fat line true false false Slovenski jezik Angleški jezik Magistrsko delo/naloga 2026-09-03 07:17:44 2026-09-03 07:17:52 2026-09-04 12:42:40 0000-00-00 00:00:00 2026 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 286704 290059779 60710.pdf 60710.pdf 1 97C693D8B815CE564E42D099976E9429 d2d1514ad9ad28c384d74f7acb5e162306cc407fb13d9bda7229fa64eb213e1c 612ee11c-a756-11f1-9b0d-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=245814 Biotehniška fakulteta 0 0 0