Vpliv krožnih RNA na izražanje genov cirkadianega ritma v celični liniji U2OS

Resnik, Katja (author), Režen, Tadeja (mentor)

Krožne molekule RNA (circRNA) so enoverižne, kovalentno sklenjene ribonukleinske kisline, ki jih uvrščamo v skupino nekodirajočih molekul RNA. Porast števila raziskav na področju krožnih molekul RNA vodi v odkrivanje čedalje več pomembnih funkcij in potencialnih načinov uporabe krožnih molekul RNA v medicini in pri procesih zdravljenja različnih bolezni. Tako se med drugim predvideva, da naj bi bile specifične krožne molekule RNA udeležene v procesih uravnavanja cirkadianega ritma, kar smo se odločili nekoliko podrobneje raziskati v sklopu naloge. Cirkadiani ritem je namreč naravno prisotno nihanje psiholoških, biokemijskih in vedenjskih procesov v organizmu, ki naj bi se periodično ponavljali približno vsakih 24 ur. Z namenom analize vpliva izbranih krožnih molekul RNA na izražanje genov cirkadianega ritma BMAL1, CLOCK, PER1, PER2, CRY1 in CRY2 je bilo v sklopu predhodnih raziskav v celicah celične linije U2OS čezmerno izraženih 11 potencialnih krožnih RNA molekul. Namen moje naloge je bil, potrditi čezmerno izraženost izbranih circRNA, nato pa za dve z največjim potencialnim vplivom na cirkadiani ritem analizirati spremembo v ravni izražanja zgoraj omenjenih genov med celicami, kjer je bila izbrana krožna RNA čezmerno izražena in celicami, ki so bile transficirane s praznim vektorjem pRR (brez vstavljenega zapisa za circRNA), oziroma netransficiranimi celicami, ki jim niso bili dodani niti vektorji, niti reagenti transfekcije. V sklopu eksperimentalnega dela smo tako uspešno potrdili čezmerno izraženost izbranih krožnih molekul RNA v celicah celične linije U2OS, prav tako pa smo uspeli zaznati spremembo v ravni izražanja gena BMAL1 v primeru čezmerne izraženosti circRNA hsa_circ_0001380 in gena PER2 v primeru čezmerne izraženosti circRNA hsa_circ_0000524. Odkrili pa smo tudi vpliv samega praznega vektorja pRR na izražanje genov PER1 in BMAL1.

• Language: Slovenian
• Keywords: krožne RNA molekule, hsa_circ_0000524, hsa_circ_0001380, cirkadiani ritem
• Work type: Bachelor thesis/paper
• Typology: 2.11 - Undergraduate Thesis
• Organization: FKKT - Faculty of Chemistry and Chemical Technology
• Year: 2023
• PID: 20.500.12556/RUL-151981
• COBISS.SI-ID: 174529539
• Publication date in RUL: 26.10.2023

RESNIK, Katja, 2023, Vpliv krožnih RNA na izražanje genov cirkadianega ritma v celični liniji U2OS [online]. Bachelor’s thesis. [Accessed 23 April 2025]. Retrieved from: https://repozitorij.uni-lj.si/IzpisGradiva.php?lang=eng&id=151981

Secondary language

• Language: English
• Title: Impact of circular RNAs on the expression of circadian rhythm genes in the U2OS cell line

Abstract

Circular RNA molecules (circRNA) are single-stranded, covalently closed ribonucleic acids that belong to the group of non-coding RNA molecules. The increasing number of studies on circular RNA molecules has led to the discovery of more and more important functions and potential uses of circular RNA molecules in medicine and in the treatment processes of various diseases. It is also hypothesized that specific circular RNA molecules are involved in regulating circadian rhythm, which we have decided to investigate in more detail as part of thesis. Circadian rhythm is a naturally occurring fluctuation of psychological, biochemical, and behavioral processes in the body that is believed to repeat periodically every 24 hours. In order to analyze the impact of selected circular RNA molecules on the expression of circadian rhythm genes BMAL1, CLOCK, PER1, PER2, CRY1, and CRY2, 11 potential circular RNA molecules were overexpressed in U2OS cell line cells as part of previous research. The purpose of my thesis was to confirm the overexpression of the selected circRNAs and then analyze the changes in the expression levels of the aforementioned genes between cells where the circular RNA was overexpressed, cells transfected with an empty vector pRR (without an inserted circRNA sequence), and non-transfected cells to which neither vectors nor transfection reagents were added. In the experimental part, we successfully confirmed the overexpression of the selected circular RNA molecules in U2OS cell line cells, and we also detected changes in the expression level of the BMAL1 gene in the case of overexpression of circRNA hsa_circ_0001380 and the PER2 gene in the case of overexpression of circRNA hsa_circ_0000524. We also discovered the influence of the empty vector pRR itself on the expression of the PER1 and BMAL1 genes.

• Keywords: circular RNA molecules, has_circ_0000524, has_circ_0001380, circadian rhythm