Department of Chemical Engineering, University of Seoul, 163 Seoulsiripdaero, Dongdaemungu, Seoul 02504, Republic of Korea.
ACS Appl Mater Interfaces. 2020 Dec 16;12(50):55554-55563. doi: 10.1021/acsami.0c12506. Epub 2020 Dec 1.
Advances in the DNA nanotechnology have enabled the fabrication of DNA-based hydrogels with precisely controlled structures and tunable mechanical and biological properties. Compared to DNA hydrogel, preparation of RNA-based hydrogel remains challenging due to the inherent instability of naked RNA. To overcome these limitations, we fabricated a DNA-RNA hybrid hydrogel via stepwise dual enzymatic polymerization. Multimeric short hairpin RNAs (shRNAs) were hybridized with functional DNA aptamers for targeting and mechanical properties of the hydrogel. The obtained DNA-RNA hybrid hydrogel was ultrasoft, robust, and injectable hence reconfigurable into any confined structures. As a model system, the hydrogel was able to mimic microtubule structures under physiological conditions and designed to release the functional small interfering RNA (siRNA)-aptamer complex (SAC) sequentially. In addition, we encoded restriction enzyme-responsive sites in DNA-RNA hybrid hydrogel to boost the release of SAC. This novel strategy provides an excellent platform for systematic RNA delivery through double-controlled release, SAC release from hydrogel, and subsequent release of siRNA from the SAC, which has promising potential in RNA therapy.
DNA 纳米技术的进步使得基于 DNA 的水凝胶的制造成为可能，这种水凝胶具有精确控制的结构和可调的机械和生物特性。与 DNA 水凝胶相比，由于裸露 RNA 的固有不稳定性，制备基于 RNA 的水凝胶仍然具有挑战性。为了克服这些限制，我们通过逐步双重酶聚合制备了 DNA-RNA 杂合水凝胶。多聚短发夹 RNA (shRNA) 与功能 DNA 适体杂交，以赋予水凝胶靶向性和机械性能。所得的 DNA-RNA 杂合水凝胶超柔软、坚固且可注射，因此可重新配置成任何受限结构。作为模型系统，该水凝胶能够在生理条件下模拟微管结构，并设计为顺序释放功能小干扰 RNA (siRNA)-适体复合物 (SAC)。此外，我们在 DNA-RNA 杂合水凝胶中编码了限制酶响应性位点，以促进 SAC 的释放。这种新策略通过双控制释放、从水凝胶中释放 SAC 以及随后从 SAC 中释放 siRNA 为系统的 RNA 递供提供了一个极好的平台，在 RNA 治疗方面具有广阔的应用前景。
