Llanos Roxana M, Ke Bi-Xia, Wright Magali, Deal Yolanda, Monty Francois, Kramer David R, Mercer Julian F B
Centre for Cellular and Molecular Biology, School of Life and Environmetal Sciences, Deakin University, Burwood 3125, Australia.
Biochim Biophys Acta. 2006 Apr;1762(4):485-93. doi: 10.1016/j.bbadis.2005.12.011. Epub 2006 Feb 3.
The brindled mouse is an accurate model of the fatal human X-linked copper deficiency disorder, Menkes disease. Males carrying the mutant allele of the Menkes gene orthologue Atp7a die in the second week of life. To determine whether the genetic defect in the brindled mice could be corrected by expression of the human Menkes gene, male transgenic mice expressing ATP7A from the chicken beta-actin composite promoter (CAG) were mated with female carriers of the brindled mutation (Atp7a(Mo-br)). Mutant males carrying the transgene survived and were fertile but the copper defect was not completely corrected. Unexpectedly males corrected with one transgenic line (T25#5) were mottled and resembled carrier females, this effect appeared to be caused by mosaic expression of the transgene. In contrast, males corrected with another line (T22#2) had agouti coats. Copper concentrations in tissues of the rescued mutants also resembled those of the heterozygous females, with high levels in kidney (84.6+/-4.9 microg/g in corrected males vs. 137.0+/-44.3 microg/g in heterozygotes) and small intestine (15.6+/-2.5 microg/g in corrected males vs. 15.7+/-2.8 microg/g in heterozygotes). The results show that the Menkes defect in mice is corrected by the human Menkes gene and that adequate correction is obtained even when the transgene expression does not match that of the endogenous gene.
斑驳小鼠是人类致命的X连锁铜缺乏症——门克斯病的精确模型。携带门克斯基因直系同源物Atp7a突变等位基因的雄性小鼠在出生后第二周死亡。为了确定人类门克斯基因的表达是否能纠正斑驳小鼠的基因缺陷,将从鸡β-肌动蛋白复合启动子(CAG)表达ATP7A的雄性转基因小鼠与携带斑驳突变(Atp7a(Mo-br))的雌性携带者交配。携带转基因的突变雄性小鼠存活且可育,但铜缺陷并未完全纠正。出乎意料的是,用一个转基因系(T25#5)纠正的雄性小鼠出现斑驳毛色,类似于携带者雌性小鼠,这种效应似乎是由转基因的嵌合表达引起的。相比之下,用另一个系(T22#2)纠正的雄性小鼠具有野生型毛色。获救突变体组织中的铜浓度也与杂合子雌性小鼠相似,肾脏中铜含量较高(纠正后的雄性小鼠为84.6±4.9微克/克,杂合子为137.0±44.3微克/克),小肠中铜含量也较高(纠正后的雄性小鼠为15.6±2.5微克/克,杂合子为15.7±2.8微克/克)。结果表明,人类门克斯基因可纠正小鼠的门克斯缺陷,即使转基因表达与内源基因表达不匹配,也能实现充分纠正。