Department of Chemistry, University of North Carolina at Charlotte, Charlotte, NC, USA.
Department of Biological Sciences, University of North Carolina at Charlotte, Charlotte, NC, USA.
Methods Mol Biol. 2023;2709:211-228. doi: 10.1007/978-1-0716-3417-2_14.
Nucleic acid nanoparticles (NANPs) are extensively investigated as diagnostic and therapeutic tools. These innovative particles can be composed of RNA, DNA, and/or modified nucleic acids. Due to the regulatory role of nucleic acids in the cellular system, NANPs have the ability to identify target molecules and regulate expression of genes in disease pathways. However, translation of NANPs in clinical settings is hindered due to inefficient intracellular delivery, chemical instability, and off-target immunostimulatory effects following immune recognition. The composition of nucleic acids forming NANPs has been demonstrated to influence immunorecognition, subcellular compartmentalization, and physicochemical properties of NANPs. This chapter first outlines the methods used to generate a panel of NANPs with a uniform shape, size, charge, sequence, and connectivity. This includes the procedures for replacing the RNA strands with DNA or chemical analogs in the designated NANPs. Second, this chapter will also describe experiments to assess the effect of the chemical modification on enzymatic and thermodynamic stability, delivery efficiency, and subcellular compartmentalization of NANPs.
核酸纳米颗粒 (NANPs) 被广泛研究作为诊断和治疗工具。这些创新的颗粒可以由 RNA、DNA 和/或修饰的核酸组成。由于核酸在细胞系统中的调节作用，NANPs 能够识别靶分子并调节疾病途径中基因的表达。然而，由于细胞内递送效率低下、化学不稳定性以及免疫识别后的非靶向免疫刺激性作用，NANPs 在临床环境中的转化受到阻碍。形成 NANPs 的核酸的组成已被证明会影响免疫识别、亚细胞区室化和 NANPs 的物理化学性质。本章首先概述了生成一组具有均匀形状、大小、电荷、序列和连接性的 NANPs 的方法。这包括用 DNA 或化学类似物替换指定 NANPs 中的 RNA 链的程序。其次，本章还将描述评估化学修饰对酶和热力学稳定性、递送效率和 NANPs 亚细胞区室化的影响的实验。
