Kloniranje smernih in protismernih ponovitev GGGGCC v vektor pRINT in njihova detekcija v sesalskih celičnih kulturah

Ivošević, Vanja (author), Župunski, Vera (mentor)

Pri bolezni amiotrofična lateralna skleroza je najpogostejša mutacija povečanje števila ponovitev GGGGCC tudi do tisočkrat v intronu gena C9orf72. V plazmid pRINT se vnese zapis za smerne in protismerne ponovitve GGGGCC. Plazmid zapisuje rdeči fluorescenčni protein. V intron se lahko vnese zapis za željeni protein in tako nastane reporterski sistem, ki dokazuje, da se zapis za željeni protein (v tem primeru ponovitve GGGGCC) res prepisuje. Nastane mRNA in intron, v katerem so ponovitve ali zapis za željeni protein, se izreže. Delovanje reporterskega sistema se potrdi tudi z detekcijo ponovitev v celicah.

• Language: Slovenian
• Keywords: amiotrofična lateralna skleroza, heksanukleotidne ponovitve GGGGCC, vektor pRINT
• 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-148945
• COBISS.SI-ID: 165233667
• Publication date in RUL: 01.09.2023

Secondary language

• Language: English
• Title: Cloning of sense and antisense GGGGCC repeats into the pRINT vector and their detection in mammalian cell cultures

Abstract

The most common mutation detected in amyotrophic lateral sclerosis is an increase in the number of GGGGCC repeats up to a thousand times in the intron of the C9ORF72 gene. We will insert sense and antisense GGGGCC repeats into the intron of red fluorescence protein in the pRINT plasmid. This way a reporter system is formed, which confirms that GGGGCC repeats are transcribed if the cells are red.

• Keywords: amyotrophic lateral sclerosis, hexanucleotide repeats GGGGCC, pRINT vector