A Mouse Strain Where Basal Connective Tissue Growth Factor Gene Expression Can Be Switched from Low to High
A Mouse Strain Where Basal Connective Tissue Growth Factor Gene Expression Can Be Switched from Low to High
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DOI：
10.1371/journal.pone.0012909
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发表时间：
2010-09-22
期刊：
影响因子：
3.7
通讯作者：
Maeda, Nobuyo
中科院分区：
文献类型：
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作者：
Doherty, Heather E.;Kim, Hyung-Suk;Maeda, Nobuyo
结缔组织生长因子（CTGF）是一种信号分子，主要在细胞外基质的维持和修复中发挥作用。Ctgf表达增加与慢性器官损伤中的纤维化相关。然而，研究CTGF在体内纤维化疾病中的作用受到Ctgf缺失小鼠的围产期致死率以及先前Ctgf过度产生的小鼠模型的有限范围的阻碍。在这里，我们设计了一种新的方法，并设计了一个单一的突变小鼠品系，其中内源性Ctgf-3 '非翻译区（3'UTR）被替换为含有两个串联排列的3'UTR序列的盒。修饰的Ctgf等位基因使用来自小鼠FBJ骨肉瘤癌基因（c-Fos）的3'UTR，并产生不稳定的mRNA，导致60%的正常Ctgf表达（Lo等位基因）。在Cre表达后，切除c-Fos-3'UTR产生利用更稳定的牛生长激素（bGH）3'UTR的转录物，导致Ctgf表达增加（Hi等位基因）。使用Ctgf Lo和Hi突变体，并与Ctgf敲除或Cre表达小鼠杂交，我们已经产生了一系列具有30倍范围Ctgf表达的菌株。Ctgf表达最低的小鼠，正常的30%，看起来很健康，而Ctgf的9倍过度表达导致异常，包括发育迟缓和颅面缺陷，以及E10 - 12的胚胎死亡。另一方面，在出生后的生活中，由他莫昔芬诱导的Cre过度表达Ctgf与生活相容。Ctgf Lo-Hi突变小鼠应证明有助于进一步了解CTGF在纤维化疾病中的功能。此外，该方法可用于产生在其他基因中具有定量变异的小鼠品系，特别是具有广泛表达的基因、在不同组织中具有不同功能的基因、或其中改变的基因表达与正常发育不相容的基因。
Connective tissue growth factor (CTGF) is a signaling molecule that primarily functions in extracellular matrix maintenance and repair. Increased Ctgf expression is associated with fibrosis in chronic organ injury. Studying the role of CTGF in fibrotic disease in vivo, however, has been hampered by perinatal lethality of the Ctgf null mice as well as the limited scope of previous mouse models of Ctgf overproduction. Here, we devised a new approach and engineered a single mutant mouse strain where the endogenous Ctgf-3' untranslated region (3'UTR) was replaced with a cassette containing two 3'UTR sequences arranged in tandem. The modified Ctgf allele uses a 3'UTR from the mouse FBJ osteosarcoma oncogene (c-Fos) and produces an unstable mRNA, resulting in 60% of normal Ctgf expression (Lo allele). Upon Cre-expression, excision of the c-Fos-3'UTR creates a transcript utilizing the more stable bovine growth hormone (bGH) 3'UTR, resulting in increased Ctgf expression (Hi allele). Using the Ctgf Lo and Hi mutants, and crosses to a Ctgf knockout or Cre-expressing mice, we have generated a series of strains with a 30-fold range of Ctgf expression. Mice with the lowest Ctgf expression, 30% of normal, appear healthy, while a global nine-fold overexpression of Ctgf causes abnormalities, including developmental delay and craniofacial defects, and embryonic death at E10-12. Overexpression of Ctgf by tamoxifen-inducible Cre in the postnatal life, on the other hand, is compatible with life. The Ctgf Lo-Hi mutant mice should prove useful in further understanding the function of CTGF in fibrotic diseases. Additionally, this method can be used for the production of mouse lines with quantitative variations in other genes, particularly with genes that are broadly expressed, have distinct functions in different tissues, or where altered gene expression is not compatible with normal development.