Faithful tissue-specific expression of the human chromosome 21-linked COL6A1 gene in BAC-transgenic mice.
Faithful tissue-specific expression of the human chromosome 21-linked COL6A1 gene in BAC-transgenic mice.
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人类 21 号染色体连接的 COL6A1 基因在 BAC 转基因小鼠中忠实地组织特异性表达。
DOI：
10.1007/s00335-006-0082-y
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发表时间：
2007
期刊：
影响因子：
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通讯作者：
Tycko,Benjamin
中科院分区：
文献类型：
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作者：
Xing,Luzhou;Salas,Martha;Lin,Chyuan-Sheng;Zigman,Warren;Silverman,Wayne;Subramaniyam,Shivakumar;Murty,VundavalliV;Tycko,Benjamin
我们用含有人类col6a1基因的细菌人工染色体（BAC）建立了转基因小鼠。在高拷贝和低拷贝转基因系中，我们发现col6a1mrna在时间和空间上的正确表达，与小鼠的9个成人器官和4个胎儿器官中col6a1mrna的表达相似。唯一的例外是胎儿肺，与内源基因相比，转基因在胎儿肺中的表达较差。Western blotting和免疫组织化学显示，转基因col6a1mrna的表达与拷贝数相关，基因剂量的增加与皮肤和心脏中胶原VI α 1的产生增加相关。col6a1位于21号染色体上，该基因已成为唐氏综合症患者心脏缺陷和皮肤异常的候选基因。低拷贝和高拷贝的col6a1转基因基因以正常的孟德尔比例出生和存活，没有心脏畸形或皮肤组织学改变。这些数据表明，在这个167kb的BAC克隆中存在调节col6a1表达的主要启动子和增强子序列。col6a1bac转基因小鼠缺乏强烈的心脏或皮肤表型，这表明该基因表达的增加本身并不能解释唐氏综合征的这些表型。
We created transgenic mice with a bacterial artificial chromosome (BAC) containing the humanCOL6A1gene. In high-copy and low-copy transgenic lines, we found correct temporal and spatial expression ofCOL6A1mRNA, paralleling the expression of the murineCol6a1gene in a panel of nine adult and four fetal organs. The only exception was the fetal lung, in which the transgene was expressed poorly compared with the endogenous gene. Expression ofCOL6A1mRNA from the transgene was copy number-dependent, and the increased gene dosage correlated with increased production of collagen VI alpha 1 in skin and heart, as indicated by Western blotting and immunohistochemistry.COL6A1maps to Chromosome 21 and this gene has been a candidate for contributing to cardiac defects and skin abnormalities in Down syndrome. The low-copy and high-copyCOL6A1transgenics were born and survived in normal Mendelian proportions, without cardiac malformations or altered skin histology. These data indicate that the major promoter and enhancer sequences regulatingCOL6A1expression are present in this 167-kb BAC clone. The lack of a strong cardiac or skin phenotype in theCOL6A1BAC-transgenic mice suggests that the increased expression of this gene does not, by itself, account for these phenotypes in Down syndrome.