苯硼酸修饰的聚乙烯亚胺系统在 T 细胞小 RNA 转染的应用
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国家科技部港澳台合作专项项目(2015DFH50230);国家自然科学基金项目(31400746);深圳市基础研究自由探索项目 (JCYJ20160531171724757);上海市卫计委项目(201440428);上海市长宁区科委重大项目(cnkw2014z06)

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Delivery of Small RNAs by Phenylboronic Acid-Grafted Polyethylenimine Nanoparticles into T Lymphocytes
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    摘要:

    小 RNA 是 T 细胞发育、分化和功能的重要调控分子,但目前仍缺乏安全有效的 T 细胞 RNA 转 染系统。基于 T 细胞表面唾液酸化的特性,该文利用苯硼酸和唾液酸的相互作用,带动苯硼酸修饰的聚乙烯亚胺系统(Polyethylenimine-Phenylboronic Acid,PEI-PBA)介导小 RNA 的 T 细胞转染。通过细胞计数试剂盒(CCK-8)和羟基荧光素二醋酸盐琥珀酰亚胺脂(CFSE)实验显示,PEI-PBA 转染系统对 T 细胞无显著细胞毒性和异常增殖。同时,流式检测人源抗 CD3/28 磁珠激活下的分化抗原 3(CD3)阳性 T 细胞在 PEI-PBA 转染系统下,小 RNA 平均摄取率增加到18.43%,而在鼠源 T 细胞中 PEI-PBA 介导的小 RNA 转染并无明显效果。结果显示,PEI-PBA 纳米转染系统介导小 RNA 的递送具有无毒高效的特点。

    Abstract:

    Small RNAs are essential regulators for T cell development, differentiation and functions. However, it is hard to deliver small RNAs into primary T cells by conventional transfection methods. Here, we reported that amphiphilic PBA-grafted PEI1.8k (PEI-PBA) nanovector facilitates the primary T cell-targeted RNA delivery miRNAthrough recognition of sialic groups on cell membrane of the T lymphocytes. CCK-8 (cell counting kit-8) and CFSE (5,6-carboxyfluorescein diacetate, succinimidyl ester) assay showed that the administration of PEI-PBA did not cause significant cell toxicity or abnormal proliferation. Meanwhile, the flowcytometry showed the average intake of miRNA was increased to 18.43% in the anti-CD3/CD28 activated CD3+ (cluster of differentiation 3) T lymphocytes by PEI-PBA in human, but not mouse. The results showed that PEI-PBA nano-system had effective delivery of small RNAs in human primary T cells without toxcity.

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引文格式
黎桂妃,贡一峰,金 言,等.苯硼酸修饰的聚乙烯亚胺系统在 T 细胞小 RNA 转染的应用 [J].集成技术,2018,7(1):51-58

Citing format
LI Guifei, GONG Yifeng, JIN Yan, et al. Delivery of Small RNAs by Phenylboronic Acid-Grafted Polyethylenimine Nanoparticles into T Lymphocytes[J]. Journal of Integration Technology,2018,7(1):51-58

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  • 收稿日期:2017-10-26
  • 最后修改日期:2017-12-06
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  • 在线发布日期: 2018-01-22
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