Department of Biological Systems Engineering, University of Nebraska-Lincoln, Lincoln, NE, USA.
Adjuvance Technologies Inc., Lincoln, NE, USA.
Methods Mol Biol. 2024;2720:165-176. doi: 10.1007/978-1-0716-3469-1_12.
Gene delivery via the oral route offers a promising strategy for improving DNA vaccination and gene-based therapy outcomes. The noninvasive nature of oral delivery lends to ease of dosing, which can facilitate convenience and patient compliance. Moreover, oral administration allows for both local and systemic production of therapeutic genes or, in the case of DNA vaccination, mucosal and systemic immunity. Here, we describe the methods to produce a dual biomaterial, oral DNA delivery system composed of chitosan (CS) and zein (ZN). In this system, CS serves to encapsulate and deliver DNA cargo to intestinal cells in the form of CS-DNA nanoparticles (CS-DNA NPs), while ZN is used to form a protective matrix around the CS-DNA NPs that prevent degradation during gastric transit but then degrades to release the CS-DNA NPs for transfection upon entry into the intestines. These particles have demonstrated the ability to effectively protect cargo DNA from simulated gastric degradation in vitro and mediate transgene production in vivo, making them an effective oral gene delivery system.
经口腔途径递送基因提供了一种有前途的策略，可改善 DNA 疫苗接种和基于基因的治疗效果。口服递送的非侵入性性质有利于剂量给药，这可以方便和提高患者的顺应性。此外，口服给药允许局部和全身产生治疗性基因，或者在 DNA 疫苗接种的情况下，粘膜和全身免疫。在这里，我们描述了生产双生物材料的方法，该材料由壳聚糖 (CS) 和玉米醇溶蛋白 (ZN) 组成的口服 DNA 递送系统。在该系统中，CS 用于将 DNA 货物包裹并以 CS-DNA 纳米颗粒 (CS-DNA NPs) 的形式递送至肠细胞，而 ZN 用于在 CS-DNA NPs 周围形成保护基质，以防止在胃转运过程中降解，但随后降解以在进入肠道时释放 CS-DNA NPs 进行转染。这些颗粒已证明能够有效地保护货物 DNA 免受体外模拟胃降解，并在体内介导转基因的产生，使它们成为有效的口服基因递送系统。
