北京生物醫(yī)學(xué)工程

巰基烷基化殼聚糖載綠色熒光蛋白質(zhì)粒基因納米粒子的制備與研究

Preparation and Studies of Thiolated N Alkylated Chitosan Green Fluorescent Protein pDNA Nanoparticles

作者：慕卿趙偉亮白欣李樹寶盧迪方月娥趙宇

單位：中國科學(xué)技術(shù)大學(xué)（合肥230026）

關(guān)鍵詞：殼聚糖；巰基化；烷基化；納米粒子；基因轉(zhuǎn)染；水溶性

分類號(hào)：

出版年·卷·期（頁碼）：2010·29·1（6-10）

摘要：

合成了新型的基因載體巰基烷基化殼聚糖（TACS），采用復(fù)凝聚法制備了巰基烷基化殼聚糖基因納米粒子，并表征其形態(tài)和粒徑。利用體外轉(zhuǎn)染實(shí)驗(yàn)定性評(píng)估了納米粒子的轉(zhuǎn)染效率。結(jié)果表明，粒子形態(tài)不很均一，粒徑在390nm左右；凝膠電泳實(shí)驗(yàn)證明了載體能有效地包裹和保護(hù)基因不受DNase Ⅰ酶的消化；溶解實(shí)驗(yàn)證實(shí)了巰基化增強(qiáng)了TACS的水溶性；體外基因轉(zhuǎn)染實(shí)驗(yàn)證實(shí)了TACS能夠在人胚胎腎細(xì)胞（HEK293）轉(zhuǎn)染基因，并且其轉(zhuǎn)染效率遠(yuǎn)高于殼聚糖納米粒子。這些結(jié)果表明，巰基烷基化殼聚糖有望成為有價(jià)值的基因載體。

Thiolated Nalkylated chitosan （TACS） was developed as a novel tool for gene delivery and the modified chitosanpDNA nanoparticles were prepared by complex coacervation. The morphology of nanoparticles was irregular， observed by transmission electronic microscopy （TEM）.The average particle size was 390nm determined by nanoparticle size analyzer. The TACSpDNA nanoparticles could partially protect the encapsulated plasmid pDNA from nuclease degradation as shown by electrophoretic mobility analysis. The solubility test showed sulfhedrylated reaction enhanced the water solubility of TACS. The gene transfection experiment in vitro suggested that the TACSpDNA nanoparticles could transfect HEK293 effectively and the transfection efficiency of TACSpDNA nanoparticles was much higher than that of chitosanpDNA nanoparticles. All these identified TACS as a promising vector for gene delivery.

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