|Tipo di tesi||Tesi di laurea magistrale|
|Titolo||Studio del binding tra fattore trascrizionale NF-Y e DNA attraverso un biosensore EGOFET.|
|Titolo in inglese||Investigation of the binding between transcription factor NF-Y and DNA by means of an EGOFET biosensor.|
|Struttura||Dipartimento di Scienze della Vita|
|Corso di studi||BIOTECNOLOGIE INDUSTRIALI (D.M. 270/04)|
|Data inizio appello||2021-10-14|
|Disponibilità||Embargo di 18 mesi|
|Data di rilascio||2023-04-14|
Il fattore di trascrizione nucleare Y (NF-Y) è un fattore di trascrizione trimerico legante CCAAT il cui reguloma, interattoma e potenziale oncogenico indicano un coinvolgimento diretto nella trasformazione cellulare (Dolfini et al., 2019). Dati di ricerca precedenti mostrano che la sovraespressione della sua subunità A (NF-YA) provoca una deregolamentazione di molti geni pro-crescita dipendenti da CCAAT nel BRCA (cancro al seno) e in altri tumori.
Nuclear transcription Factor Y (NF-Y) is a CCAAT-binding, trimeric transcription factor whose regulome, interactome and oncogenic potential point to direct involvement in cellular transformation. Previous research data shows that the overexpression of its A subunit (NF-YA) results in a deregulation of many CCAAT-dependent pro-growth genes in BRCA (Breast cancer) and other tumors. Moreover, NF-YA can be found in two different forms, NF-YAs (short isoform) and NF-YAl (long isoform), which could be associated with different progression stage of the tumor. Biosensors are devices capable of performing a sensitive and selective detection of a wide range of targets (such as molecules, proteins, oligonucleotides up to virus and whole cells), thanks to their synergy with biological probes which are naturally specialized, or can be engineered, for a specific target. In this work we used Electrolye-Gated Field-Effect Transistors (EGOFET) where the biorecognition process takes place on an electrode where the whole system is located, and that interacts with an organic semiconductor-based transistor. Due to the changes in electrical properties associated to the recognition phenomenon, the modulation operated by the sensing electrode on the semiconductor is affected, resulting in a measurable variation of the transduced signal. A TIPS-Pentacene based, top gated EGOFET device was realized to perform the detection of NF-Y transcription factor in its trimeric form, by functionalizing the gate electrode with a self-assembled monolayer and immobilizing onto that a double-strand DNA oligonucleotide containing the CCAAT sequence. Detection of the DNA sequence itself was also performed, by immobilizing a single strand onto the device and incubating in the complementary single strand oligonucleotide instead of NF-Y. Main goal of the study is to realize the first EGOFET biosensor for the detection of a transcription factor, namely NF-Y trimer with the long isoform of subunit A. This research could be a valid starting point for the creation of a powerful tool for both in vitro DNA-TF interaction studies and clinical analysis in tumor diagnostics and characterization, by discrimination and quantification of the two NF-YA isoforms.