用于在小鼠中进行cre介导的条件性敲除的Smad5基因floxed等位基因的构建。

Generation of a floxed allele of Smad5 for cre-mediated conditional knockout in the mouse.

作者信息

Umans Lieve, Vermeire Liesbeth, Francis Annick, Chang Hua, Huylebroeck Danny, Zwijsen An

机构信息

Department of Developmental Biology (VIB 7), Flanders Interuniversity Institute for Biotechnology (VIB), and Laboratory of Molecular Biology (CELGEN), University of Leuven, Leuven, Belgium.

出版信息

Genesis. 2003 Sep;37(1):5-11. doi: 10.1002/gene.10219.

DOI:10.1002/gene.10219

PMID:14502571

Abstract

Smad5 is a member of the Smad family of intracellular mediators of BMP signals and in endothelial cells of TGF-beta signals. We and others previously showed that loss of Smad5 in the mouse results in embryonic lethality (between E9.5-E11.5) due to multiple embryonic and extraembryonic defects. To circumvent the early embryonic lethality and to allow tissue- and time-specific Smad5 inactivation, we created a conditional Smad5 allele in the mouse. Floxed Smad5 (Smad5(flE2,Neo/flE2,Neo)) mice were generated in which both exon2 and the Neo-cassette were flanked by loxP sites. Here we demonstrate that embryos with ubiquitous Cre-mediated deletion of Smad5 (Smad5(flDeltaE2/flDeltaE2)) phenocopy the conventional Smad5 knockout mice. Smad5(flE2/flE2) mice are now available and will be a valuable tool to analyze the role of Smad5 beyond its crucial early embryonic function throughout development and postnatal life.

摘要

Smad5是骨形态发生蛋白（BMP）信号以及转化生长因子-β（TGF-β）信号在细胞内的介质Smad家族的成员。我们和其他人之前表明，小鼠中Smad5缺失会导致胚胎致死（在E9.5-E11.5之间），原因是多种胚胎和胚外缺陷。为了规避早期胚胎致死性并实现组织和时间特异性的Smad5失活，我们在小鼠中创建了一个条件性Smad5等位基因。构建了floxed Smad5（Smad5(flE2,Neo/flE2,Neo)）小鼠，其中外显子2和新霉素盒两侧均有loxP位点。在此我们证明，通过Cre介导的Smad5普遍缺失的胚胎（Smad5(flDeltaE2/flDeltaE2)）与传统的Smad5基因敲除小鼠表现出相似的表型。现在已有Smad5(flE2/flE2)小鼠，它将成为一个有价值的工具，用于分析Smad5在整个发育过程和出生后生活中除关键的早期胚胎功能之外的作用。