Gambari Roberto, Gasparello Jessica, Fabbri Enrica, Borgatti Monica, Tamanini Anna, Finotti Alessia
Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
Department of Molecular Pathology and Diagnostics, University Hospital of Verona, Verona, Italy.
Methods Mol Biol. 2020;2105:199-215. doi: 10.1007/978-1-0716-0243-0_12.
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.
微小RNA参与人类疾病已得到确凿证实。因此，对微小RNA活性进行药理学调控似乎是开发新型药物（微小RNA疗法）的一个非常有趣的方法。一个重要的研究领域是在罕见病领域开发微小RNA疗法的可能性。在这方面，引人关注的分子基于肽核酸（PNA），在首次描述时，其显示出由N-（2-氨基乙基）甘氨酸单元组成的假肽主链，并且被发现是反义疗法和反基因疗法的优秀候选物。本文的目的是描述用于确定旨在靶向参与囊性纤维化的微小RNA的肽核酸活性的方法，使用miR-145-5p及其靶标囊性纤维化跨膜传导调节因子（CFTR）mRNA作为模型系统。通过讨论使用人肺上皮Calu-3细胞系作为细胞模型系统获得的数据，介绍了用于研究靶向miR-145-5p的肽核酸作用的方法。
