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在通过饮用水提供荧光素后对活体转基因小鼠中的基因表达进行成像。

Imaging gene expression in live transgenic mice after providing luciferin in drinking water.

作者信息

Hiler Daniel J, Greenwald Megan L, Geusz Michael E

机构信息

Department of Biological Sciences and the J. P. Scott Center of Neuroscience, Mind, and Behavior, Bowling Green State University, Bowling Green, Ohio 43403-0212, USA.

出版信息

Photochem Photobiol Sci. 2006 Nov;5(11):1082-5. doi: 10.1039/b608360a. Epub 2006 Aug 25.

Abstract

Mice expressing the firefly luciferase gene luc under the control of various gene promoters are used to image long-term changes in tumor growth, infection, development, and circadian rhythms. This novel approach enables ongoing regulation of gene expression to be visualized through repeated imaging of luciferase bioluminescence. Typically, luciferin, the luciferase substrate, is injected into mice before they are anaesthetized for imaging. To avoid the effects of handling and stress from injection on expression of the transgene, oral luciferin delivery methods were tested as an alternative to current methods. For unobscured imaging, a transgenic mouse line containing luc controlled by the enhancer and promoter for the major immediate-early gene of human cytomegalovirus (CMV) was crossed with a hairless albino mouse stock (HRS/J), resulting in the Hr-CMV line. Mice given food and water ad libitum readily drank 1-5 mM luciferin in water or apple juice and could be imaged repeatedly on subsequent days without any apparent adverse effects. Oral and injected luciferin produced similar patterns of luminescence in the body areas examined: abdomen, tail vertebrae, gonads, hind leg, foreleg and others, although the tail showed a slightly brighter relative luminescence after oral luciferin. These results show that luciferin is not appreciably degraded in the digestive tract and can be easily administered orally to avoid injection and any concomitant effects on behavior that could alter gene expression.

摘要

在各种基因启动子控制下表达萤火虫荧光素酶基因luc的小鼠,被用于对肿瘤生长、感染、发育和昼夜节律的长期变化进行成像。这种新方法能够通过对荧光素酶生物发光的重复成像来可视化基因表达的持续调控。通常,在对小鼠进行麻醉以进行成像之前,会向其注射荧光素酶的底物荧光素。为了避免处理和注射应激对转基因表达的影响,人们测试了口服荧光素递送方法作为当前方法的替代方案。为了获得清晰的成像效果,将一个含有由人类巨细胞病毒(CMV)主要立即早期基因的增强子和启动子控制的luc的转基因小鼠品系与一个无毛白化小鼠品系(HRS/J)杂交,得到了Hr-CMV品系。随意进食和饮水的小鼠很容易饮用含1-5 mM荧光素的水或苹果汁,并且在随后的几天里可以反复成像,没有任何明显的不良影响。口服和注射荧光素在检查的身体部位(腹部、尾椎、性腺、后腿、前腿等)产生了相似的发光模式,不过口服荧光素后尾巴的相对发光略亮一些。这些结果表明,荧光素在消化道中不会明显降解,并且可以很容易地通过口服给药,以避免注射以及任何可能改变基因表达的伴随行为影响。

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