A series of ENU-induced single-base substitutions in a long-range cis-element altering Sonic hedgehog expression in the developing mouse limb bud
A series of ENU-induced single-base substitutions in a long-range cis-element altering Sonic hedgehog expression in the developing mouse limb bud
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DOI：
10.1016/j.ygeno.2006.09.005
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发表时间：
2007-02-01
期刊：
影响因子：
4.4
通讯作者：
Shiroishi, Toshihiko
中科院分区：
文献类型：
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作者：
Masuya, Hiroshi;Sezutsu, Hideki;Shiroishi, Toshihiko
哺乳动物-鱼-保守序列1(MFCS1)是一个高度保守的序列，作为Sonic hedgehog(Shh)表达的肢体特异性顺式作用调节因子，位于小鼠Shh编码序列的1Mb之外。通过对ENU致突变小鼠档案的基因驱动筛选，我们获得了在MFCS1中有三个新的点突变的小鼠：M101116、M10111 7和M101192。表型分析显示，M101116小鼠在肢芽前缘表现出轴前多指和异位Shh表达，就像先前发现的突变M100081一样。相比之下，M10111 7和M101192在肢体形态上未见明显异常。此外，转基因分析表明，除了正常的后部表达外，M101116和MI00081序列还驱动了翼芽前缘的异位报告基因的表达。在携带由M101117驱动的报告基因TransgeDe的胚胎中没有观察到这种异位表达。这些结果表明，M101116和M100081影响MFCSI的负性调节活性，从而抑制发育中的肢芽中前Shh的表达。因此，这项研究表明，基因驱动的ENU诱导突变的筛选是探索保守的非编码序列和潜在的顺式调控元件功能的有效方法。(C)2006 Elsevier Inc.保留所有权利。
Mammal-fish-conserved-sequence 1 (MFCS1) is a highly conserved sequence that acts as a limb-specific cis-acting regulator of Sonic hedgehog (Shh) expression, residing 1 Mb away from the Shh coding sequence in mouse. Using gene-driven screening of an ENU-mutagenized mouse archive, we obtained mice with three new point mutations in MFCS 1: M101116, M10111 7, and M101192. Phenotype analysis revealed that M101116 mice exhibit preaxial polydactyly and ectopic Shh expression at the anterior margin of the limb buds like a previously identified mutant, M100081. In contrast, M10111 7 and M101192 show no marked abnon-nalities in limb morphology. Furthermore, transgenic analysis revealed that the M101116 and MI00081 sequences drive ectopic reporter gene expression at the anterior margin of the limb bud, in addition to the normal posterior expression. Such ectopic expression was not observed in the embryos carrying a reporter transgeDe driven by M101117. These results suggest that M101116 and M100081 affect the negative regulatory activity of MFCSI, which suppresses anterior Shh expression in developing limb buds. Thus, this study shows that gene-driven screening for ENU-induced mutations is an effective approach for exploring the function of conserved, noncoding sequences and potential cis-regulatory elements. (c) 2006 Elsevier Inc. All rights reserved.