明膠/DNA 奈米微粒於基因轉殖雞之應用

Abstract

本實驗將利用明膠當作為一個新的基因載體，並做出粒徑為300nm左右的明膠奈米微粒。且於DNA的包覆率測試上，發現其包覆率可達到80％以上；佐以雷射奈米粒徑暨界面電位測量儀、穿透式電子顯微鏡證實此明膠奈米微粒的粒徑與穩定性。且經由DNase Ι的作用反應可以說明明膠奈米微粒具有保護DNA的能力。於HeLa細胞的轉染實驗中，發現明膠奈米微粒和市售商品LipofectaineTM 2000有幾乎相似的轉染效率，但是相對於市售商品LipofectaineTM 2000，明膠奈米微粒有較高的細胞活性和較低的細胞毒性。於轉染雞胚的實驗中，經由明膠奈米微粒轉染的雞胚其存活率約為80-100％，觀察被轉染四天後的雞胚也可以發現綠色螢光蛋白的產生。這說明了明膠奈米微粒的生物相容性的優勢在轉殖動物上是極具潛力的。

In thus study, we tried to use gelatin as carriers in gene delivery, and prepare the gelatin nanoparticles with particle size of 300nm. For calculating the capturing ratio, the DNA loading efficiency is nearly 80％. Size analysis and stability were performed under dynamic light scattering and TEM. DNA encapsulated in the gelatin nanoparticles could be protected from DNaseΙdegradation. In HeLa cell transfection experiments, efficiencies of commercial product LipofectaineTM 2000 and gelatin/ pEGFP-C1 nanoparticles were nearly the same. When comparing to commercial product LipofectaineTM 2000, gelatin/pEGFP-C1 nanoparticles had higher cell viability and lower cytotoxicity. After gelatin nanoparticles transfection, the egg hatchability was 80-100％. We could also detect Green fluorescence in the fourth day chicken embryo transfected by gelatin/pEGFP-C1 nanoparticles.