Structure-activity relationships of cationic shell-crosslinked knedel-like nanoparticles: shell composition and transfection efficiency/cytotoxicity.
Structure-activity relationships of cationic shell-crosslinked knedel-like nanoparticles: shell composition and transfection efficiency/cytotoxicity.
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DOI：
10.1016/j.biomaterials.2009.10.033
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发表时间：
2010-03
期刊：
影响因子：
14
通讯作者：
Wooley, Karen L.
中科院分区：
文献类型：
--
作者：
Zhang, Ke;Fang, Huafeng;Wang, Zhenghui;Li, Zhou;Taylor, John-Stephen A.;Wooley, Karen L.
阳离子纳米粒子是一类很有前途的寡核苷酸和基因传递的转染剂，但其有效性和细胞毒性差异很大。最近，我们开发了一类基于阳离子壳交联纳米颗粒（cSCK）的新型阳离子转染剂，可以用寡核苷酸和质粒DNA有效地转染哺乳动物细胞。为了在不增加细胞毒性的情况下进一步提高转染效率，我们研究了这些纳米颗粒的壳中伯胺（pa）、叔胺（ta）和羧酸（ca）基团的组成的影响。在4：1至40：1的宽范围的胺与磷酸盐比率（N/P比率）下制备cSCKs与质粒DNA（pDNA）或硫代磷酸酯2′-OMe寡核苷酸（ps-MeON）的一系列离散复合物。发现复合物的尺寸和纳米颗粒完全结合ODN的能力取决于cSCK胺与DNA磷酸盐（N/P）的比率和cSCK缓冲能力。然后通过监测来自编码的EGFP的荧光来评估cSCK用质粒DNA转染细胞的能力，并通过监测来自ps-MeON介导的剪接校正产生的荧光素酶的发光来评估cSCK递送ps-MeON的能力。然而，发现阳离子cSCK-pa 25-ta 75以20：1的N/P比最适合于将质粒DNA转染到HeLa细胞中，而cSCK-pa 50-ta 50以10：1的N/P比最适合于ps-MeON递送。我们还发现，增加叔胺相对于伯胺的比例降低了细胞毒性。这些结果表明，通过将叔胺掺入cSCKs的壳中，可以实现基因和寡核苷酸递送效率的显著改善以及HeLa细胞中细胞毒性的降低。
Cationic nanoparticles are a promising class of transfection agents for oligonucleotide and gene delivery, but vary greatly in their effectiveness and cytotoxicity. Recently, we developed a new class of cationic transfection agents based on cationic shell-crosslinked nanoparticles (cSCKs) that efficiently transfect mammalian cells with both oligonucleotides and plasmid DNA. In an effort to further improve transfection efficiency without increasing cytotoxicity, we examined the effects of the composition of primary amine (pa), tertiary amine (ta) and carboxylic acid (ca) groups in the shell of these nanoparticles. A series of discrete complexes of the cSCKs with plasmid DNA (pDNA) or phosphorothioate 2′-OMe oliogonucleotides (ps-MeON) were prepared over a broad range of amine to phosphate ratios (N/P ratio) of 4:1 to 40:1. The sizes of the complexes and the ability of the nanoparticles to completely bind ODNs were found to depend on the cSCK amine to DNA phosphate (N/P) ratio and the cSCK buffering capacity. The cSCKs were then evaluated for their ability to transfect cells with plasmid DNA by monitoring fluorescence from an encoded EGFP, and for delivery of ps-MeON by monitoring luminescence from luciferase resulting from ps-MeON-mediated splicing correction. Whereas the cationic cSCK-pa25-ta75 was found to be best for transfecting plasmid DNA into HeLa cells at an N/P ratio of 20:1, cSCK-pa50-ta50, at an N/P ratio 10:1 was best for ps-MeON delivery. We also found that increasing the proportion of tertiary relative to primary amine reduced the cytotoxicity. These results demonstrate that a dramatic improvement in gene and oligonucleotide delivery efficiency with decreased cytotoxicity in HeLa cells can be achieved by incorporation of tertiary amines into the shells of cSCKs.
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影响因子：
6.2
作者：
Sun, Guorong;Hagooly, Aviv;Welch, Michael J.
通讯作者：
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影响因子：
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作者：
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影响因子：
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作者：
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通讯作者：
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DOI：
10.1002/1099-0518(200012)38:1
发表时间：
2000-01-01
影响因子：
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作者：
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通讯作者：
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影响因子：
4.9
作者：
Fang, Huafeng;Zhang, Ke;Taylor, John-Stephen A.
通讯作者：
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