Efficient Gene Transfection by Histidine-Modified Chitosan through Enhancement of Endosomal Escape
Efficient Gene Transfection by Histidine-Modified Chitosan through Enhancement of Endosomal Escape
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DOI：
10.1021/bc1000609
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发表时间：
2010-06-01
影响因子：
4.7
通讯作者：
Hashida, Mitsuru
中科院分区：
文献类型：
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作者：
Chang, Kai-Ling;Higuchi, Yuriko;Hashida, Mitsuru
壳聚糖具有作为生物相容性基因载体的潜力。然而，壳聚糖的转染效率低，因为缓慢的内体逃逸速率。组氨酸在内体范围内的缓冲能力将有助于质粒DNA（pDNA）从内体逃逸。本研究在壳聚糖中引入组氨酸以提高转染效率。壳聚糖和组氨酸通过2-亚氨基硫烷和半胱氨酸提供的二硫键连接。通过将壳聚糖或组氨酸修饰的壳聚糖与质粒DNA混合制备复合物。组氨酸修饰的壳聚糖具有更宽的缓冲范围，且组氨酸修饰的壳聚糖/pDNA复合物的细胞摄取率高于壳聚糖/pDNA复合物。壳聚糖/四甲基罗丹明（TMR）-pDNA复合物在胞液中被捕获在囊泡中，而组氨酸修饰的壳聚糖携带的TMR-pDNA在胞液中分布更广。这一结果表明，组氨酸可以在高缓冲能力的帮助下帮助pDNA从内体逃逸。组氨酸修饰的壳聚糖/pDNA复合物的基因表达高于壳聚糖/pDNA复合物。这些结果表明组氨酸修饰提高了壳聚糖的转染效率。
Chitosan has the potential to be a biocompatible gene carrier. However, the transfection efficiency of chitosan is low because of the slow endosomal escape rate. The buffering capacity of histidine in the endosomal range would help the escape of plasmid DNA (pDNA) from endosomes. In this study, histidine was introduced into chitosan to improve the transfection efficiency. Chitosan and histidine were linked by disulfide bonds provided by 2-iminothiolane and cysteine. The complexes were prepared by mixing chitosan or histidine-modified chitosan with plasmid DNA. A broader buffering range of histidine-modified chitosan was observed, and the cellular uptake of histidine-modified chitosan/pDNA complexes was higher than that of chitosan/pDNA complexes. Although chitosan/tetramethylrhodamine (TMR)-pDNA complexes were trapped in the vesicles in cytosol, TMR-pDNA carried by histidine-modified chitosan was more widely distributed in the cytosol. This result suggests that histidine can help pDNA escape from endosomes with the help of the high buffering capacity. The gene expression of histidine-modified chitosan/pDNA complexes was higher than that of chitosan/pDNA complexes. These results suggest that histidine modification improves the transfection efficiency of chitosan.