Utišavanje gena Scarlet metodom RNA interferencije kod rakova porodice Asellidae (Crustacea: Isopoda)

master's thesis

• Title (english): Silencing of the Scarlet gene using RNA interference in asellid crustaceans (Crustacea: Isopoda: Asellidae)
• Author: Martina Kuzman
• Mentor: Helena Bilandžija (mentor)
• Mentor: Domagoj Đikić (komentor)
• Committee member: Domagoj Đikić (predsjednik povjerenstva)
• Committee member: Biljana Balen (član povjerenstva)
• Committee member: Dubravko Pavoković (član povjerenstva)
• Committee member: Maja Matulić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2024-09-23, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

RNA interferencija je metoda kojom se, unosom dsRNA u organizam, uzrokuje utišavanje gena degradacijom ciljne mRNA. Na rakovima je to sve učestalija metoda istraživanja funkcije gena te je najbolja metoda unosa dsRNA mikroinjektiranje. Jednakonožni slatkovodni rakovi vrste Asellus aquaticus i Proasellus coxalis pokazuju iznimnu fenotipsku plastičnost pigmentacije tijela. U pigmentaciju tijela jednakonožnih rakova su uključeni omokromi; molekule koje nastaju od aminokiseline triptofana. U put biosinteze omokroma su uključeni enzimi i ABC transporteri, jedan od kojih je i protein Scarlet. U ovom istraživanju sam uz pomoć RNA interferencije, mikroinjektiranjem dsRNA gena Scarlet u organizam navedenih životinja pokušala utišati gen Scarlet i na taj način uzrokovati fenotipsku promjenu smanjenja pigmentacije tijela. Kod vrste A. aquaticus mikroinjektiranjem dsRNA gena Scarlet za vrijeme presvlačenja došlo je do značajnog smanjenja pigmentacije tijela. Mikroinjektirane jedinke vrste P. coxalis imale su visoku smrtnost, ali je primijećen trend posvijetljenja pigmentacije tijela. Najbolji efekt primijećen je kod jedinki mikroinjektiranih za vrijeme presvlačenja kod obje vrste.

Abstract (english)

RNA interference is a method used for silencing specific genes by introducing dsRNA into the organism which causes degradation of the target mRNA. This method is increasingly used for understanding the functions of genes in Crustaceans, and it was shown that the best method of introducing the dsRNA is by microinjection. Isopods Asellus aquaticus and Proasellus coxalis show high levels of phenotypic plasticity in body pigmentation. Ommochromes, molecules biosynthesized from the amino acid tryptophan, are responsible for body pigmentation in isopods. The ommochrome biosynthesis involves different enzymes and ABC transporters, including the protein Scarlet. Here, I tried to silence the Scarlet gene using RNA interference by microinjecting dsRNA in the organisms of the two above mentioned species. I expected this would cause a change in body pigmentation which would be shown as brightening of body color. In A. aquaticus microinjecting the dsRNA of the Scarlet gene in animals which were in moulting showed a significant reduction in body pigmentation. Microinjected P. coxalis showed high levels of mortality, but a trend of reduction in body pigmentation was noticed. The best effect was shown in animals which were in moulting at the time of microinjecting.

• Keywords: dsRNA; omokromi; Asellus aquaticus; Proasellus coxalis; mikroinjektiranje
• Keywords (english): dsRNA; ommochromes; Asellus aquaticus; Proasellus coxalis; microinjection
• Language: croatian
• URN:NBN: urn:nbn:hr:217:361224
• Project: Number: TTP-2018-07-9675; Title: Evolucija u tami; Title: Evolution in the dark; Acronym: Evodark; Leader: Helena Bilandžija; Jurisdiction: Croatia; Funder: Hrvatska zaklada za znanost; Funding stream: Tenure Track Pilot Programme
• Study programme: Title: Experimental Biology-University graduate programme; Study programme type: university; Study level: graduate; Academic / professional title: sveučilišni/a magistar/magistra eksperimentalne biologije (sveučilišni/a magistar/magistra eksperimentalne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2025-02-11 14:40:27