The rat probasin gene promoter directs hormonally and developmentally regulated expression of a heterologous gene specifically to the prostate in transgenic mice.
The rat probasin gene promoter directs hormonally and developmentally regulated expression of a heterologous gene specifically to the prostate in transgenic mice.
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DOI：
10.1210/mend.8.2.8170479
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发表时间：
1994-02
影响因子：
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通讯作者：
N. Greenberg;F. Demayo;P. Sheppard;Roberto J. Barrios;R. Lebovitz;M. Finegold;R. Angelopoulou;R. Angelopoulou;J. Dodd;M. L. Duckworth;J. Rosen;R. Matusik
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文献类型：
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作者：
N. Greenberg;F. Demayo;P. Sheppard;Roberto J. Barrios;R. Lebovitz;M. Finegold;R. Angelopoulou;R. Angelopoulou;J. Dodd;M. L. Duckworth;J. Rosen;R. Matusik
携带426个碱基的大鼠前列腺素(PB)基因启动子和28个碱基的5‘-非翻译区的表达盒足以靶向细菌氯霉素乙酰转移酶(CAT)基因在转基因小鼠的前列腺中的表达。PB-CAT转基因在五个独立品系中的三个(60%)小鼠中表达，并且这种表达，正如先前报道的内源性大鼠基因一样，是男性特有的，主要局限于前列腺的外侧、背侧和腹叶，只有在前列腺前和精囊中检测到非常低水平的CAT活性。转基因的发育和激素调节也与大鼠基因的报道相似，在2-7周龄的小鼠前列腺中观察到CAT活性增加了70倍，这一时间对应于性成熟。去势后前列腺组织中PB-CAT活性下降至去势前水平的3.5%，补充睾酮后，去势后雄性小鼠前列腺组织中的CAT活性接近去势前水平。地塞米松不能诱导去势雄鼠的转基因表达。将PB-CAT与鸡溶菌酶基因基质附着区共注射，可使PB-CAT与鸡溶菌酶基因基质结合区共整合，进一步将PB-CAT的表达模式限制在前列腺背外侧，并抑制其在前列腺腹侧的表达。这些研究表明，一个最小的大鼠前盆启动子可以以发育和激素调节的方式针对前列腺特异性的异源基因表达。
An expression cassette carrying 426 basepairs of the rat probasin (PB) gene promoter and 28 basepairs of 5'-untranslated region is sufficient to target the expression of the bacterial chloramphenicol acetyltransferase (CAT) gene specifically to the prostate in transgenic mice. The PB-CAT transgene was expressed in three of five (60%) independent lines of mice, and this expression, as reported previously for the endogenous rat gene, was male specific, restricted primarily to the lateral, dorsal, and ventral lobes of the prostate, with only very low levels of CAT activity detected in the anterior prostate and seminal vesicles. The developmental and hormonal regulation of the transgene also paralleled that reported for the rat gene, with a 70-fold increase in CAT activity in the mouse prostate observed between 2-7 weeks of age, a time corresponding to sexual maturation. PB-CAT activity in the prostate declined after castration to 3.5% of the precastration level, and the CAT activity in castrated males approached precastration levels when mice were supplemented with testosterone. Transgene expression in castrated males was not induced by dexamethasone. Coinjection of PB-CAT with a chicken lysozyme gene matrix attachment region resulted in their cointegration and further restricted the pattern of PB-CAT to the dorsolateral prostate, with suppressed expression observed in the ventral prostate. These studies demonstrate that a minimal rat probasin promoter can target heterologous gene expression specifically to the prostate in a developmentally and hormonally regulated fashion.