Development of Polymer–Lipid Hybrid Nanoparticles for Large-Sized Plasmid DNA Transfection

转染 纳米颗粒 聚合物 质粒 DNA 纳米技术 材料科学 生物 基因 生物化学 复合材料
作者
Masatoshi Maeki,Shuya Uno,Kaisei Sugiura,Yusuke Sato,Yoichiro Fujioka,Akihiko Ishida,Yusuke Ohba,Hideyoshi Harashima,Manabu Tokeshi
出处
期刊:ACS Applied Materials & Interfaces [American Chemical Society]
卷期号:16 (2): 2110-2119 被引量:7
标识
DOI:10.1021/acsami.3c14714
摘要

RNA and DNA delivery technologies using lipid nanoparticles (LNPs) have advanced significantly, as demonstrated by their successful application in mRNA vaccines. To date, commercially available RNA therapeutics include Onpattro, a 21 bp siRNA, and mRNA vaccines comprising 4300 nucleotides for COVID-19. However, a significant challenge remains in achieving efficient transfection, as the size of the delivered RNA and DNA increases. In contrast to RNA transfection, plasmid DNA (pDNA) transfection requires multiple steps, including cellular uptake, endosomal escape, nuclear translocation, transcription, and translation. The low transfection efficiency of large pDNA is a critical limitation in the development of artificial cells and their cellular functionalization. Here, we introduce polymer-lipid hybrid nanoparticles designed for efficient, large-sized pDNA transfection. We demonstrated that LNPs loaded with positively charged pDNA-polycation core nanoparticles exhibited a 4-fold increase in transfection efficiency for 15 kbp pDNA compared with conventional LNPs, which encapsulate a negatively charged pDNA-polycation core. Based on assessments of the size and internal structure of the polymer-lipid nanoparticles as well as hemolysis and cellular uptake analysis, we propose a strategy to enhance large-sized pDNA transfection using LNPs. This approach holds promise for accelerating the in vivo delivery of large-sized pDNA and advancing the development of artificial cells.
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