Conditional gene expression in the respiratory epithelium of the mouse
Conditional gene expression in the respiratory epithelium of the mouse
复制标题
DOI：
10.1023/a:1013986627504
复制
发表时间：
2002-02-01
影响因子：
3
通讯作者：
Whitsett, JA
中科院分区：
文献类型：
--
作者：
Perl, AKT;Tichelaar, JW;Whitsett, JA
利用SP-C和CCSP启动子控制下表达的反向四环素反式激活因子（rtTA）开发了介导呼吸道上皮基因表达的条件性时空调控的转基因小鼠模型。荧光素酶活性在胎儿和成年双转基因小鼠的肺中检测到，但在其他组织或单转基因小鼠中未检测到。在成年小鼠中，检测到最大荧光素酶活性后16小时，在饮用水中的多西环素，或2小时后注射多西环素。在食物颗粒中给予多西环素后观察到转基因的激活。在长时间暴露于多西环素后，去除多西环素后荧光素酶活性缓慢下降，表明维持转基因表达的组织库的重要性。在SP-C-rtTA小鼠中，妊娠母鼠暴露于多西环素诱导早在E10.5胎鼠肺组织中的荧光素酶活性。荧光素酶活性维持在哺乳期期间，当母亲接受强力霉素的饮用水中的幼鼠的肺组织。在CCSP-rtTA小鼠中，在不存在多西环素的情况下未检测到荧光素酶。在SP-C-rtTA小鼠中，在不存在多西环素的情况下检测到荧光素酶活性，但通过施用药物增强约10倍。SP-C-rtTA和CCSP-rtTA激活剂小鼠控制发育和成熟呼吸道上皮中转基因的表达，将有助于研究肺中的基因功能。
Transgenic mouse models mediating conditional temporal and spatial regulation of gene expression to the respiratory epithelium were developed utilizing the reverse tetracycline transactivator (rtTA) expressed under the control of SP-C and CCSP promoters. Luciferase activity was detected in the lungs of fetal and adult double transgenic mice but was not detected in other tissues or in single transgenic mice. In adult mice, maximal luciferase activity was detected 16 h after the administration of doxycycline in the drinking water, or 2 h after the injection of doxycycline. Activation of the transgene was observed after the administration of doxycycline in food pellets. After prolonged exposure to doxycycline, luciferase activity decreased slowly following removal of doxycycline, suggesting the importance of tissue pools which maintained expression of the transgene. In SP-C-rtTA mice, exposure of the pregnant dam to doxycycline induced luciferase activity in fetal lung tissue as early as E10.5. Luciferase activity was maintained in the lung tissue of pups during the period of lactation when the mother received doxycycline in the drinking water. In the CCSP-rtTA mice, luciferase was not detected in the absence of doxycycline. In the SP-C-rtTA mice, luciferase activity was detected in the absence of doxycycline but was enhanced approximately 10-fold by administration of drugs. The SP-C-rtTA and CCSP-rtTA activator mice control the expression of transgenes in the developing and mature respiratory epithelium, and will be useful for the study of gene function in the lung.