Hartmann Alyssa K, Cairns-Gibson Dominic F, Santiana Joshua J, Tolentino Mark Q, Barber Halle M, Rouge Jessica L
Department of Chemistry, University of Connecticut, Storrs, CT, 06269, USA.
Chembiochem. 2018 Jun 3. doi: 10.1002/cbic.201800302.
Herein, we describe the characterization of a novel self-assembling and intracellular disassembling nanomaterial for nucleic acid delivery and targeted gene knockdown. By using a recently developed nucleic acid nanocapsule (NAN) formed from surfactants and conjugated DNAzyme (DNz) ligands, it is shown that DNz-NAN can enable cellular uptake of the DNAzyme and result in 60 % knockdown of a target gene without the use of transfection agents. The DNAzyme also exhibits activity without chemical modification, which we attribute to the underlying nanocapsule design and release of hydrophobically modified nucleic acids as a result of enzymatically triggered disassembly of the NAN. Fluorescence-based experiments indicate that the surfactant-conjugated DNAzymes are better able to access a fluorescent mRNA target within a mock lipid bilayer system than the free DNAzyme, highlighting the advantage of the hydrophobic surfactant modification to the nucleic acid ligands. In vitro characterization of DNz-NAN's substrate-cleavage kinetics, stability in biological serum, and persistence of knockdown against a proinflammatory transcription factor, GATA-3, are presented.
在此，我们描述了一种用于核酸递送和靶向基因敲低的新型自组装和细胞内拆解纳米材料的特性。通过使用由表面活性剂和共轭脱氧核酶（DNz）配体形成的最近开发的核酸纳米胶囊（NAN），结果表明，DNz-NAN能够使细胞摄取脱氧核酶，并在不使用转染剂的情况下导致靶基因敲低60%。脱氧核酶在未经化学修饰的情况下也表现出活性，我们将其归因于潜在的纳米胶囊设计以及由于NAN的酶促触发拆解而释放的疏水修饰核酸。基于荧光的实验表明，与游离脱氧核酶相比，表面活性剂共轭脱氧核酶在模拟脂质双层系统中能够更好地接近荧光mRNA靶标，突出了疏水表面活性剂修饰对核酸配体的优势。本文还介绍了DNz-NAN的底物切割动力学、在生物血清中的稳定性以及对促炎转录因子GATA-3的敲低持久性的体外特性。
