Using inositol as a biocompatible ligand for efficient transgene expression.
Using inositol as a biocompatible ligand for efficient transgene expression.
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使用肌醇作为生物相容性配体进行高效转基因表达
DOI：
10.2147/ijn.s77002
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发表时间：
2015
影响因子：
8
通讯作者：
Wu Y
中科院分区：
文献类型：
--
作者：
Zhang L;Bellis SL;Fan Y;Wu Y
使用阳离子聚合物如聚乙烯亚胺（PEI）和PEI衍生物作为基因载体的转基因转染技术已经显示出功效，尽管它们也具有缺点。PEI具有良好的DNA结合能力和良好的细胞内化性能，但它们不能非常有效地将基因有效载荷递送到细胞核。在这项研究中，三个超支化聚甘油-聚乙烯亚胺（PG 6-PEI）聚合物与肌醇（INO）分子的共轭发展。所得的三种PG 6-PEI-INO聚合物每个分子的INO配体数量增加。PG 6-PEI-INO 1每分子仅具有14个羧甲基INO（CMINO）单元。PG 6-PEI-INO 2每分子具有约130个CMINO单元。PG 6-PEI-INO 3具有高达约415个CMINO单位。将PG 6-PEI-INO聚合物与DNA混合产生紧凑的纳米复合材料。然后，我们使用荧光显微镜进行定位研究。随着PG 6-PEI-INO聚合物中缀合肌醇配体的数量增加，293 T细胞核内聚合物的积累相应增加。当使用PG 6-PEI-INO 3作为载体时，用球形293 T细胞进行的转染产生82%的EGFP阳性细胞。研究进一步揭示，与未与肌醇缀合的PEI和PG 6-PEI相比，细胞外三磷酸腺苷（eATP）可以抑制PG 6-PEI-INO聚合物的转基因效率。我们的工作揭示了使用肌醇作为转基因表达的有效配体的可能性。
Transgene transfection techniques using cationic polymers such as polyethylenimines (PEIs) and PEI derivatives as gene vectors have shown efficacy, although they also have shortcomings. PEIs have decent DNA-binding capability and good cell internalization performance, but they cannot deliver gene payloads very efficiently to cell nuclei. In this study, three hyperbranched polyglycerol-polyethylenimine (PG6-PEI) polymers conjugated with myo-inositol (INO) molecules were developed. The three resulting PG6-PEI-INO polymers have an increased number of INO ligands per molecule. PG6-PEI-INO 1 had only 14 carboxymethyl INO (CMINO) units per molecule. PG6-PEI-INO 2 had approximately 130 CMINO units per molecule. PG6-PEI-INO 3 had as high as 415 CMINO units approximately. Mixing PG6-PEI-INO polymers with DNA produced compact nanocomposites. We then performed localization studies using fluorescent microscopy. As the number of conjugated inositol ligands increased in PG6-PEI-INO polymers, there was a corresponding increase in accumulation of the polymers within 293T cell nuclei. Transfection performed with spherical 293T cells yielded 82% of EGFP-positive cells when using PG6-PEI-INO 3 as the vehicle. Studies further revealed that extracellular adenosine triphosphate (eATP) can inhibit the transgene efficiency of PG6-PEI-INO polymers, as compared with PEI and PG6-PEI that were not conjugated with inositol. Our work unveiled the possibility of using inositol as an effective ligand for transgene expression.