Organic and Biomimetic Chemistry Research Group, Ghent University, Krijgslaan 281 S4, B-9000 Ghent, Belgium.
Department of Chemistry, Life Sciences and Environmental Sustainability, University of Parma, Parco Area delle Scienze, 17/A, I-43124 Parma, Italy.
Methods. 2023 Oct;218:210-223. doi: 10.1016/j.ymeth.2023.08.010. Epub 2023 Aug 19.
While natural oligonucleotides (ONs) are increasingly used as therapeutic and diagnostic tools, they still face certain challenges such as low resistance to enzymatic degradation, potential immunogenicity, and delivery issues, which can limit their applications. Peptide Nucleic Acids (PNAs) are promising alternatives due to their high affinity for DNA and RNA, the high resistance to enzymatic degradation, and the easy introduction of a wide range of potential modifications. Chemical modifications that enable the covalent targeting of specific DNA and RNA strands offer additional advantages, including enhanced potency. The current study focuses on the utilization of furan-PNAs as pro-reactive probe systems and their applications to DNA and RNA targeting. Specifically, in this methodological paper, we provide practical insights into the design, synthesis, and application of furan-containing PNA probes for achieving efficient PNA-DNA and PNA-RNA interstrand crosslinking (ICL), as well as ON-templated PNA-PNA ligation systems. Furthermore, we discuss the applications of these probes in targeting DNA secondary structures, such as G-quadruplexes and i-motifs, target pull-down assays, and on-surface detection.
虽然天然寡核苷酸 (ONs) 越来越多地被用作治疗和诊断工具，但它们仍然面临一些挑战，例如对酶降解的抵抗力低、潜在的免疫原性和递送问题，这可能限制了它们的应用。肽核酸 (PNA) 是一种很有前途的替代品，因为它们与 DNA 和 RNA 的亲和力高，对酶的降解有很高的抵抗力，并且可以很容易地引入广泛的潜在修饰。能够共价靶向特定 DNA 和 RNA 链的化学修饰提供了额外的优势，包括增强的效力。本研究重点研究了呋喃-PNA 作为前反应性探针系统的利用及其在 DNA 和 RNA 靶向中的应用。具体来说，在本方法学论文中，我们提供了关于设计、合成和应用含呋喃 PNA 探针的实用见解，以实现高效的 PNA-DNA 和 PNA-RNA 链间交联 (ICL) 以及 ON 模板化 PNA-PNA 连接系统。此外，我们讨论了这些探针在靶向 DNA 二级结构（如 G-四联体和 i-基序）、靶标下拉测定和表面检测中的应用。
