Williams R S, Johnston S A, Riedy M, DeVit M J, McElligott S G, Sanford J C
Duke University, Durham, NC 27706.
Proc Natl Acad Sci U S A. 1991 Apr 1;88(7):2726-30. doi: 10.1073/pnas.88.7.2726.
Foreign genes were expressed in liver and skin cells of live mice by using a new apparatus to accelerate DNA-coated microprojectiles into tissues. After introduction of a plasmid in which the firefly luciferase gene was controlled by the human beta-actin promoter, luciferase activity was detectable for up to 14 days in mouse tissues (skin and liver). In situ hybridization histochemistry revealed that microprojectiles penetrated through multiple cell layers without evidence of tissue injury and that 10-20% of the cells in the bombarded area expressed the foreign gene. An advantage of the new design is that internal organs, such as liver, can be transfected without subjecting the tissue to a vacuum. This procedure potentially is applicable to a wide variety of tissues and cell types for studies of transcriptional control elements and for expression of foreign proteins in intact animals.
通过使用一种新装置将包裹有DNA的微弹加速进入组织,可使外源基因在活小鼠的肝脏和皮肤细胞中表达。导入一种质粒(其中萤火虫荧光素酶基因由人β-肌动蛋白启动子控制)后,在小鼠组织(皮肤和肝脏)中可检测到长达14天的荧光素酶活性。原位杂交组织化学显示,微弹穿透多个细胞层,且无组织损伤迹象,并且在轰击区域10%-20%的细胞表达了外源基因。新设计的一个优点是,诸如肝脏等内部器官无需在真空中处理组织即可进行转染。该方法有可能适用于多种组织和细胞类型,用于研究转录控制元件以及在完整动物中表达外源蛋白。