Manson A L, Trezise A E, MacVinish L J, Kasschau K D, Birchall N, Episkopou V, Vassaux G, Evans M J, Colledge W H, Cuthbert A W, Huxley C
Imperial College School of Medicine at St Mary's, London, UK.
EMBO J. 1997 Jul 16;16(14):4238-49. doi: 10.1093/emboj/16.14.4238.
We have made transgenic mice carrying a 320 kb YAC with the intact human cystic fibrosis transmembrane regulator (CFTR) gene. Mice that only express the human transgene were obtained by breeding with Cambridge null CF mice. One line has approximately two copies of the intact YAC. Mice carrying this transgene and expressing no mouse cftr appear normal and breed well, in marked contrast to the null mice, where 50% die by approximately 5 days after birth. The chloride secretory responses in these mice are as large or larger than in wild-type tissues. Expression of the transgene is highly cell type specific and matches that of the endogenous mouse gene in the crypt epithelia throughout the gut and in salivary gland tissue. However, there is no transgene expression in some tissues, such as the Brunner's glands, where it would be expected. Where there are differences between the mouse and human pattern of expression, the transgene follows the mouse pattern. We have thus defined a cloned fragment of DNA which directs physiological levels of expression in many of the specific cells where CFTR is normally expressed.
我们已经培育出携带一个含有完整人类囊性纤维化跨膜传导调节因子(CFTR)基因的320 kb酵母人工染色体(YAC)的转基因小鼠。通过与剑桥CF基因敲除小鼠杂交,获得了仅表达人类转基因的小鼠。其中一个品系大约有两份完整的YAC。携带这种转基因且不表达小鼠CFTR的小鼠看起来正常且繁殖良好,这与基因敲除小鼠形成了显著对比,在基因敲除小鼠中,约50%在出生后5天左右死亡。这些小鼠的氯离子分泌反应与野生型组织中的一样大或更大。转基因的表达具有高度的细胞类型特异性,并且与整个肠道隐窝上皮和唾液腺组织中内源性小鼠基因的表达相匹配。然而,在一些预期会有表达的组织中,如十二指肠腺,却没有转基因表达。当小鼠和人类的表达模式存在差异时,转基因遵循小鼠的模式。因此,我们已经确定了一段DNA克隆片段,它能在许多正常表达CFTR的特定细胞中指导生理水平的表达。