Hormonal modulation of a gene injected into rat heart in vivo.
Hormonal modulation of a gene injected into rat heart in vivo.
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体内注入大鼠心脏的基因的激素调节。
DOI：
10.1073/pnas.88.10.4138
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发表时间：
1991
影响因子：
11.1
通讯作者：
Leinwand,LA
中科院分区：
文献类型：
--
作者：
Kitsis,RN;Buttrick,PM;McNally,EM;Kaplan,ML;Leinwand,LA
我们通过直接注射质粒DNA的方法在体内证明了基因转移到大鼠心脏中。注射由逆转录病毒和细胞启动子驱动的基因构建体导致可检测到的报告基因活性水平。细胞启动子和5‘侧翼序列(-613-+32位)来源于大鼠α-肌球蛋白重链基因，该基因在体内的表达仅限于心肌，并受甲状腺激素的正向调节。DNA注射后，与肌球蛋白重链启动子和调控序列偶联的萤火虫荧光素酶基因的活性在心脏中被检测到，但在骨骼肌中未被检测到，并且随着甲状腺激素的处理而显著增加。因此，注射基因的表达可以针对体内特定的细胞类型，并可以受到动物荷尔蒙状态的调节。这种方法提供了一种绘制基因元件图的方法，这些基因元件调节它们对复杂刺激的反应，而这些刺激不能在体外模拟。
We demonstrate gene transfer into rat heart in vivo by the direct injection of plasmid DNA. Injection of gene constructs driven by retroviral and cellular promoters resulted in detectable levels of reporter gene activities. The cellular promoter and 5' flanking sequence (positions -613 to +32) were derived from the rat alpha-myosin heavy chain gene whose expression in vivo is restricted to cardiac muscle and is positively regulated by thyroid hormone. After DNA injection, activity of the firefly luciferase gene coupled to the myosin heavy chain promoter and regulatory sequence was detected in heart but not in skeletal muscle and was significantly increased in response to thyroid hormone treatment. Consequently, expression of injected genes can be targeted to specific cell types in vivo and can be modulated by the hormonal status of the animal. This approach provides a means of mapping the elements of genes that regulate their responses to complex stimuli that cannot be modeled in vitro.