Eswaran Lakshmanan, Kazimirsky Gila, Byk Gerardo
Laboratory of Nanobiotechnology, Department of Chemistry, Bar-Ilan University, Ramat Gan 52900, Israel.
Pharmaceutics. 2023 Jan 19;15(2):332. doi: 10.3390/pharmaceutics15020332.
The advent of protein expression using m-RNA applied lately for treating the COVID pandemic, and gene editing using CRISPR/Cas9 technology for introducing DNA sequences at a specific site in the genome, are milestones for the urgent need of developing new nucleic acid delivery systems with improved delivery properties especially for in vivo applications. We have designed, synthesized, and characterized novel cross-linked monodispersed nanohydrogels (NHG's) with well-defined sizes ranging between 50-400 nm. The synthesis exploits the formation of self-assemblies generated upon heating a thermo-responsive mixture of monomers. Self-assemblies are formed and polymerized at high temperatures resulting in NHGs with sizes that are predetermined by the sizes of the intermediate self-assemblies. The obtained NHGs were chemically reduced to lead particles with highly positive zeta potential and low cell toxicity. The NHGs form complexes with DNA, and at optimal charge ratio the size of the complexes is concomitant with the size of the NHG's. Thus, the DNA is fully embedded inside the NHGs. The new NHGs and their DNA complexes are devoid of cell toxicity which together with their tunned sizes, make them potential tools for gene delivery and foreign protein expression.
最近用于治疗新冠疫情的基于mRNA的蛋白质表达技术的出现，以及利用CRISPR/Cas9技术在基因组特定位点引入DNA序列的基因编辑技术，是迫切需要开发具有改进递送特性的新型核酸递送系统（尤其是用于体内应用）的里程碑。我们设计、合成并表征了新型交联单分散纳米水凝胶（NHG），其尺寸明确，在50至400纳米之间。合成过程利用了加热单体的热响应混合物时产生的自组装体的形成。自组装体在高温下形成并聚合，产生的NHG尺寸由中间自组装体的尺寸预先确定。所获得的NHG经过化学还原，形成具有高正zeta电位和低细胞毒性的铅颗粒。NHG与DNA形成复合物，在最佳电荷比下，复合物的尺寸与NHG的尺寸相当。因此，DNA完全嵌入NHG内部。新的NHG及其DNA复合物没有细胞毒性，再加上其调整后的尺寸，使其成为基因递送和外源蛋白表达的潜在工具。
