Issue 21, 2024

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

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.

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

Supplementary files

Article information

Article type
Paper
Submitted
18 Janv. 2024
Accepted
30 Apr. 2024
First published
30 Apr. 2024

J. Mater. Chem. C, 2024,12, 7596-7604

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

M. Sensi, A. Ricci, G. Rigillo, A. Paradisi, M. Berto, N. Gnesutta, C. Imbriano, F. Biscarini and C. A. Bortolotti, J. Mater. Chem. C, 2024, 12, 7596 DOI: 10.1039/D4TC00260A

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