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Peptide Nucleic Acids for MicroRNA Targeting

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Peptide Nucleic Acids

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2105))

Abstract

The involvement of microRNAs in human pathologies is firmly established. Accordingly, the pharmacological modulation of microRNA activity appears to be a very interesting approach in the development of new types of drugs (miRNA therapeutics). One important research area is the possible development of miRNA therapeutics in the field of rare diseases. In this respect, appealing molecules are based on peptide nucleic acids (PNAs), displaying, in their first description, a pseudo-peptide backbone composed of N-(2-aminoethyl)glycine units, and found to be excellent candidates for antisense and antigene therapies. The aim of the present article is to describe methods for determining the activity of PNAs designed to target microRNAs involved in cystic fibrosis, using as model system miR-145-5p and its target cystic fibrosis transmembrane conductance regulator (CFTR) mRNA. The methods employed to study the effects of PNAs targeting miR-145-5p are presented here by discussing data obtained using as cellular model system the human lung epithelial Calu-3 cell line.

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Abbreviations

3′UTR:

3′-Untranslated region

CF:

Cystic fibrosis

CFTR:

Cystic fibrosis transmembrane conductance regulator

EDTA:

Ethylenediaminetetraacetic acid

FBS:

Fetal bovine serum

HRP:

Horseradish peroxidase

miRNA:

MicroRNA

PBS:

Phosphate-buffered saline

PNA:

Peptide nucleic acid

RISC:

RNA-induced silencing complex

RT-qPCR:

Reverse transcription-quantitative polymerase chain reaction

SDS:

Sodium dodecyl sulfate

TBS:

Tris-buffered saline

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Acknowledgments

This work was supported by Fondazione Fibrosi Cistica (FFC), Project “Revealing the microRNAs-transcription factors network in cystic fibrosis: from microRNA therapeutics to precision medicine (CF-miRNA-THER)” FFC#7/2018. 

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Authors and Affiliations

  1. Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy

    Roberto Gambari, Jessica Gasparello, Enrica Fabbri, Monica Borgatti & Alessia Finotti

  2. Department of Molecular Pathology and Diagnostics, University Hospital of Verona, Verona, Italy

    Anna Tamanini

Authors
  1. Roberto Gambari
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  2. Jessica Gasparello
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  3. Enrica Fabbri
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  4. Monica Borgatti
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  5. Anna Tamanini
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  6. Alessia Finotti
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Corresponding author

Correspondence to Roberto Gambari .

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Editors and Affiliations

  1. Department of Cellular and Molecular Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark

    Peter E. Nielsen

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Gambari, R., Gasparello, J., Fabbri, E., Borgatti, M., Tamanini, A., Finotti, A. (2020). Peptide Nucleic Acids for MicroRNA Targeting. In: Nielsen, P. (eds) Peptide Nucleic Acids. Methods in Molecular Biology, vol 2105. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0243-0_12

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  • DOI: https://doi.org/10.1007/978-1-0716-0243-0_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0242-3

  • Online ISBN: 978-1-0716-0243-0

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