Introduction of foreign genes into tissues of living mice by DNA-coated microprojectiles.
Introduction of foreign genes into tissues of living mice by DNA-coated microprojectiles.
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通过 DNA 包被的微弹将外源基因引入活体小鼠的组织中。
DOI：
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发表时间：
1991
影响因子：
11.1
通讯作者：
J. Sanford
中科院分区：
文献类型：
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作者：
R. Williams;Stephen A. JOHNSTONtt;Mark RIEDYt;Michael J. DEVITtt;G. Sandra;Mcelligott;J. Sanford
通过使用一种新的装置来加速DNA包被的微粒进入组织，外源基因在活小鼠的肝脏和皮肤细胞中表达。在引入其中萤火虫荧光素酶基因由人β-肌动蛋白启动子控制的质粒后，在小鼠组织（皮肤和肝脏）中可检测荧光素酶活性长达14天。原位杂交组织化学显示，微弹穿透多个细胞层，没有组织损伤的证据，并且在轰击区域中10-20%的细胞表达外源基因。新设计的一个优点是，内部器官，如肝脏，可以在不使组织处于真空状态的情况下进行转染。该方法可能适用于各种组织和细胞类型，用于转录控制元件的研究和在完整动物中表达外源蛋白。
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.