Unit:
Department of BiologyLibrary of the School of Science
Author:
Mitropoulou Christina-Angeliki
Supervisors info:
Χρήστος Κοντός, Επίκουρος Καθηγητής, Τμήμα Βιολογίας, Εθνικό και Καποδιστριακό Πανεπιστήμιο Αθηνών
Original Title:
Μελέτη και επαγωγή της έκφρασης θραυσμάτων RNA του 3’ άκρου του tRNACysGCA (tRF-CysGCA) σε ανθρώπινες κυτταρικές σειρές
Translated title:
Research and induction of the expression of 3’ end of tRNACysGCA (tRFCysGCA) in human cell lines
Summary:
The main subject of this thesis is a novel class of small non-coding RNAs, derived from tRNAs, known as tRNA – derived RNA fragments. tRNA derived fragments or tRFs, derive either from precursor tRNA molecules or from mature cytoplasmic tRNAs. These RNAs are generated by a precise cleavage event, which takes place on the parental tRNA. Their biogenesis is different, depending on the cleavage spot, and demands a variety of different enzymes and complexes. Their abundance varies between different cell types and tissues, but it depends on the concentration of parental tRNAs too. There are two main classes of tRNA fragments, tRNA halves (or tiRNAs) which derive from mature tRNAs only, and smaller tRNA fragments (tRFs) which derive from both mature and precursors tRNAs.
In this diploma thesis, the induction of expression of 3’ end of tRNACysGCA (tRFCysGCA) in human cell lines was implemented. The experimental procedure included the optimization of Reverse Transcription for the proper denaturation of tRFs rather than tRNAs, the successively cloning of tRNACysGCA gene in two different vectors, and the transient transfection of the recombinant vector in human cancer cells.
The results showed that the increase in expression of 3’tRFCysGCA in transfected cancer cells was not statistically significant. Although it was the first approach in a very difficult subject, the experimental procedure can be optimized in the future. This thesis can be the first step for more experiments to pursue, which can help us understand better the functions and the interactions of these molecules in both normal and pathological conditions.
Main subject category:
Science
Keywords:
tRNA, tRF, tRNA halves, tiRNAs, tRNACys,
