作为高效质粒 CRISPR (pCRISPR) 运载工具的三种智能聚合物纳米粒子的合成、表征和细胞研究

IF 1.7 Q3 MATERIALS SCIENCE, MULTIDISCIPLINARY
Roya Khodaei, Mehrdad Bayandori, Leila Mohammad Gholinia Sarpoli, Masoumeh Souri, Iraj Hasanzade, Ronak Khodaee, Sara Saeedi, Jafar Kiani, Mahdi Karimi
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引用次数: 0

摘要

目的。基因治疗是一种通过修改基因提供治疗优势的策略,CRISPR/Cas9 是最简单、最有效的基因编辑技术。适当的智能纳米载体是将这些基因编辑工具高效送入细胞的一种可行方法。方法。本研究制备了三种新型智能纳米载体,用于将 CRISPR/Cas9 导入 PC12 细胞。我们设计了一种智能多聚物,它是由交联聚乙烯亚胺(PEI ∼ 2 KD)和 pCRISPR 利用静电相互作用制成的合成氧化还原反应聚乙烯亚胺(rPEI)。然后,为了避免与血液成分发生不必要的相互作用,分别使用了透明质酸(HA)、硫酸软骨素(CS)和海藻酸(ALG)这三种带负电荷的天然多糖作为外壳。此外,还对纳米载体的 zeta 电位、尺寸分布和负载效率进行了表征。最后,对纳米载体的细胞毒性和 GFP 基因表达进行了评估。结果外覆 HA、ALG 和 CS 的纳米载体的平均尺寸分别约为 47、66.5 和 309 nm。此外,这三种纳米载体的装载效率高,细胞摄取能力强(90%),且无明显毒性。据估计,pCRISPR 的表达量高达 21%。结论这些氧化还原反应聚合物纳米载体为 CRISPR/Cas9 的传输提供了新颖高效的载体。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Synthesis, characterization, and cellular investigation of three smart polymeric nanoparticles as efficient plasmid CRISPR (pCRISPR) delivery vehicles
Purpose. Gene therapy is a strategy to provide therapeutic advantages by modifying genes, and CRISPR/Cas9 is the simplest and the most efficient gene editing technology. Appropriate smart nanocarriers are a promising way to deliver these gene editing tools into cells efficiently. Methods. In this study, three novel smart nanocarriers were prepared for CRISPR/Cas9 delivery into PC12 cells. We designed a smart polyplex using synthetic redox-responsive polyethyleneimine (rPEI) made by crosslinked PEI ∼ 2 KD and pCRISPR using electrostatic interactions. Then, to avoid unwanted interactions with blood ingredients three natural polysaccharides with negative charge, hyaluronic acid (HA), chondroitin sulfate (CS), and alginate(ALG) have been used separately as outer shells. Additionally, nanocarriers were characterized in terms of zeta potential, size distribution, and loading efficiency. Finally, the cytotoxicity of nanocarriers and GFP gene expression were evaluated. Results. The average size of the nanocarriers with outer coats of HA, ALG, and CS was around 47, 66.5, and 309 nm, respectively. Furthermore, these three nanocarriers indicated a high loading efficiency, high capacity for cellular uptake (>90%), and no significant toxicity. The pCRISPR expression amount was estimated up to 21%. Conclusion. These redox-responsive polymeric nanocarriers suggest novel and efficient carriers for CRISPR/Cas9 delivery.
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来源期刊
Advances in Natural Sciences: Nanoscience and Nanotechnology
Advances in Natural Sciences: Nanoscience and Nanotechnology NANOSCIENCE & NANOTECHNOLOGYMATERIALS SCIE-MATERIALS SCIENCE, MULTIDISCIPLINARY
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