DNA-binding miniproteins based on zinc fingers. Assessment of the interaction using nanopores

Jéssica Rodríguez, Soraya Learte-Aymamí, Jesús Mosquera, Garbiñe Celaya, David Rodríguez-Larrea, M. Eugenio Vázquez, José L. Mascareñas

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Obtaining artificial proteins that mimic the DNA binding properties of natural transcription factors could open new ways of manipulating gene expression at will. In this context it is particularly interesting to develop simple synthetic systems. Inspired by the modularity of natural transcription factors, we have designed synthetic miniproteins that combine the zinc finger module of the transcription factor GAGA and AT-hook peptide domains. These constructs are capable of binding to composite DNA sequences of up to 14 base pairs with high affinity and good selectivity. In particular, we have synthesized three different chimeras and characterized their DNA binding properties by electrophoresis and fluorescence anisotropy. We have also used, for the first time in the study of peptide-based DNA binders, nanopore force spectroscopy to obtain further data on the DNA interaction.

Original languageEnglish
Pages (from-to)4118-4123
Number of pages6
JournalChemical Science
Volume9
Issue number17
Early online date10 Apr 2018
DOIs
Publication statusPublished - 7 May 2018

Keywords

  • DNA binding
  • chimeras
  • DNA interaction
  • nanopores

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    Rodríguez, J., Learte-Aymamí, S., Mosquera, J., Celaya, G., Rodríguez-Larrea, D., Vázquez, M. E., & Mascareñas, J. L. (2018). DNA-binding miniproteins based on zinc fingers. Assessment of the interaction using nanopores. Chemical Science, 9(17), 4118-4123. https://doi.org/10.1039/c7sc05441f