Department of Chemistry, Binghamton University, Binghamton, NY, USA.
Department of Chemistry and Biochemistry, Elizabethtown College, Elizabethtown, PA, USA.
Methods Mol Biol. 2020;2105:157-172. doi: 10.1007/978-1-0716-0243-0_9.
Because of the important roles noncoding RNAs play in gene expression, their sequence-specific recognition is important for both fundamental science and the pharmaceutical industry. However, most noncoding RNAs fold in complex helical structures that are challenging problems for molecular recognition. Herein, we describe a method for sequence-specific recognition of double-stranded RNA using peptide nucleic acids (PNAs) that form triple helices in the major grove of RNA under physiologically relevant conditions. We also outline methods for solid-phase conjugation of PNA with cell-penetrating peptides and fluorescent dyes. Protocols for PNA preparation and binding studies using isothermal titration calorimetry are described in detail.
由于非编码 RNA 在基因表达中发挥着重要作用，因此它们对序列特异性的识别对于基础科学和制药行业都很重要。然而，大多数非编码 RNA 折叠成复杂的螺旋结构，这对分子识别来说是一个挑战。在此，我们描述了一种使用肽核酸 (PNA) 识别双链 RNA 的方法，该方法在生理相关条件下 PNA 在 RNA 的大沟中形成三螺旋。我们还概述了将 PNA 与穿膜肽和荧光染料固相偶联的方法。详细描述了使用等温滴定量热法进行 PNA 制备和结合研究的方案。
