Nuclear transcription factor Y (NF-Y) is a CCAAT-binding trimeric protein. The overexpression of the DNA-binding subunit A (NF-YA) results in deregulation of many CCAAT-dependent pro-growth genes in multiple tumor types. Exon 3 alternative splicing of NF-YA results in two different isoforms, NF-YAs (short) and NF-YAl (long), which can promote tumor proliferation or metastasis, respectively. In this work, we developed an electrolyte-gated organic transistor (EGOT) biosensor to study the binding of a NF-YAl-composed NF-Y complex to its consensus sequence. We show that by using the target DNA sequence as a probe, the device detects NF-Y in the range of 1 pM to 10 nM. Control experiments performed with oligonucleotide probes mutated in the consensus sequence exhibit weaker, though not fully hindered, binding to NF-Y compared to the response to unmutated DNA. This behavior confirms that the base pairs near the CCAAT-box also have a role in the transcription factor recognition. Furthermore, we contributed to the advancement of the present state of the art by demonstrating the ability of the EGOT biosensor to detect NF-Y in cell lysate, a fundamental step towards the development of point-of-care (POC) devices for the analysis of biopsies.The first electrolyte-gated organic transistor biosensor for the detection of a transcription factor (NF-Y) in buffer and cell lysate.

Investigation of transcription factor–DNA binding with electrolyte-gated organic transistors / Sensi, Matteo; Ricci, Andrea; Rigillo, Giovanna; Paradisi, Alessandro; Berto, Marcello; Gnesutta, Nerina; Imbriano, Carol; Biscarini, Fabio; Bortolotti, Carlo Augusto. - In: JOURNAL OF MATERIALS CHEMISTRY. C. - ISSN 2050-7526. - 12:(2024), pp. 7596-7604. [10.1039/d4tc00260a]

Investigation of transcription factor–DNA binding with electrolyte-gated organic transistors

Sensi, Matteo
;
Rigillo, Giovanna;Paradisi, Alessandro;Berto, Marcello;Imbriano, Carol;Biscarini, Fabio;Bortolotti, Carlo Augusto
2024

Abstract

Nuclear transcription factor Y (NF-Y) is a CCAAT-binding trimeric protein. The overexpression of the DNA-binding subunit A (NF-YA) results in deregulation of many CCAAT-dependent pro-growth genes in multiple tumor types. Exon 3 alternative splicing of NF-YA results in two different isoforms, NF-YAs (short) and NF-YAl (long), which can promote tumor proliferation or metastasis, respectively. In this work, we developed an electrolyte-gated organic transistor (EGOT) biosensor to study the binding of a NF-YAl-composed NF-Y complex to its consensus sequence. We show that by using the target DNA sequence as a probe, the device detects NF-Y in the range of 1 pM to 10 nM. Control experiments performed with oligonucleotide probes mutated in the consensus sequence exhibit weaker, though not fully hindered, binding to NF-Y compared to the response to unmutated DNA. This behavior confirms that the base pairs near the CCAAT-box also have a role in the transcription factor recognition. Furthermore, we contributed to the advancement of the present state of the art by demonstrating the ability of the EGOT biosensor to detect NF-Y in cell lysate, a fundamental step towards the development of point-of-care (POC) devices for the analysis of biopsies.The first electrolyte-gated organic transistor biosensor for the detection of a transcription factor (NF-Y) in buffer and cell lysate.
2024
30-apr-2024
12
7596
7604
Investigation of transcription factor–DNA binding with electrolyte-gated organic transistors / Sensi, Matteo; Ricci, Andrea; Rigillo, Giovanna; Paradisi, Alessandro; Berto, Marcello; Gnesutta, Nerina; Imbriano, Carol; Biscarini, Fabio; Bortolotti, Carlo Augusto. - In: JOURNAL OF MATERIALS CHEMISTRY. C. - ISSN 2050-7526. - 12:(2024), pp. 7596-7604. [10.1039/d4tc00260a]
Sensi, Matteo; Ricci, Andrea; Rigillo, Giovanna; Paradisi, Alessandro; Berto, Marcello; Gnesutta, Nerina; Imbriano, Carol; Biscarini, Fabio; Bortolotti, Carlo Augusto
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11380/1341186
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