O'Brien John, Lummis Sarah C R
Neurobiology Division, MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK.
Brain Res Brain Res Protoc. 2002 Aug;10(1):12-5. doi: 10.1016/s1385-299x(02)00175-7.
The hand-held gene gun uses a pulse of helium to fire small gold particles coated with desiccated DNA (microcarriers) at target cells. This method of biolistic transfection is becoming increasingly popular as an effective means of rapid gene delivery into mammalian tissue. Current methods of microcarrier preparation, however, are slow (up to 2 days) and can result in variations in transfection efficiency due to a number of problems including shearing of DNA, agglomeration and adhesion of gold particles. Here we describe an improved, more rapid method of microcarrier preparation. To evaluate the new procedure we have used DNA encoding yellow fluorescent protein (EYFP), a modified version of the green fluorescent protein, which we have transfected into HEK293 cells. The data show that transfection by the new method results in high levels of transfection efficiency and low variability compared to an alternative method.
手持式基因枪利用氦气脉冲将包裹有干燥DNA（微载体）的小金颗粒射向靶细胞。这种生物弹道转染方法作为一种将基因快速导入哺乳动物组织的有效手段正变得越来越流行。然而，目前微载体的制备方法很慢（长达2天），并且由于包括DNA剪切、金颗粒团聚和粘附等许多问题，可能导致转染效率的差异。在此，我们描述了一种改进的、更快速的微载体制备方法。为了评估新方法，我们使用了编码黄色荧光蛋白（EYFP）的DNA，它是绿色荧光蛋白的一种修饰形式，我们已将其转染到HEK293细胞中。数据表明，与另一种方法相比，新方法转染可产生高水平的转染效率且变异性低。
