Significant and prolonged antisense effect of a multifunctional envelope-type nano device encapsulating antisense oligodeoxynucleotide
Significant and prolonged antisense effect of a multifunctional envelope-type nano device encapsulating antisense oligodeoxynucleotide
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DOI：
10.1211/jpp.58.4.0002
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发表时间：
2006-04-01
影响因子：
3.3
通讯作者：
Harashima, H
中科院分区：
文献类型：
--
作者：
Nakamura, Y;Kogure, K;Harashima, H
以8-精氨酸(R8)为内化配体，研制了一种多功能包膜型纳米器件(MEND)，作为一种高效的非病毒载体系统用于载体DNA(PDNA)的传递。制备了三种类型的R8-Mend，将编码荧光素酶的pDNA与由硬脂酰辛基精氨酸、多L赖氨酸和鱼精蛋白三种阳离子缩合的反义寡核苷酸共包裹，并在体外分析了其反义作用。以鱼精蛋白为凝聚剂包装的ODN-Mend在16h后反义效果达90%，持续48h反义效果可达75%以上，效果明显优于LipofetAmine2000。另一方面，以PLL和STR-R8为冷凝剂制备的ODN-Mend对荧光素酶活性没有明显的抑制作用。尽管ODN-Mend的理化性质与其反义作用之间没有特定的关系，但ODN-Mend之间的反义作用模式与R8-Mend包埋的siRNA载体DNA的沉默效应相似。这些结果表明，R8-Mend能够将包裹的DNA运送到胞浆和胞核，鱼精蛋白也可以作为一种有效的解缩剂，不仅在胞核中，而且在胞浆中。综上所述，我们成功地利用鱼精蛋白作为DNA缩合剂和解缩剂开发了一种具有高度反义效果的ODN-Mend。
A multifunctional envelope-type nano device (MEND) was developed for use as an efficient non-viral system for the delivery of plasmid DNA (pDNA) using octaarginine (R8) as an internalizing ligand. Three types of R8-MENDs were prepared, co-encapsulating luciferase-encoding pDNA and antiluciferase oligodeoxynucleotide (ODN) condensed by three polycations, stearyl octaarginine (STR-R8), poly-L-lysine (PLL) and protamine, and the antisense effects of the ODN-encapsulated R8-MENDs (ODN-MEND) were analysed in-vitro. The ODN-MEND packaged using protamine as a condenser showed a 90% antisense effect 16 h after the transfection, and a persistent antisense effect of over 75% for up to 48 h, which was much more effective than that of LipofectAmine2000. On the other hand, the ODN-MENDs prepared using PLL and STR-R8 as condensers did not show any significant inhibition of luciferase activity. Although there was no specific relation between the physicochemical characteristics of the ODN-MENDs and their antisense effect, the pattern of the antisense effect among the ODN-MENDs was similar to that of the silencing effect of R8-MEND encapsulating plasmid DNA encoding siRNA. These results suggest that R8-MENDs are able to deliver encapsulated DNA to the cytosol as well as to the nucleus, and that protamine can also function as an efficient decondenser, not only in the nucleus but also in the cytosol. In conclusion, we successfully developed an ODN-MEND with a high antisense effect using protamine as a DNA condensing as well as a decondensing agent.