Hemizygous Le-Cre transgenic mice have severe eye abnormalities on some genetic backgrounds in the absence of LoxP sites.
Hemizygous Le-Cre transgenic mice have severe eye abnormalities on some genetic backgrounds in the absence of LoxP sites.
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DOI：
10.1371/journal.pone.0109193
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
West JD
中科院分区：
文献类型：
--
作者：
Dorà NJ;Collinson JM;Hill RE;West JD
在Le-CreTg/−; Pax 6 fl/+小鼠中研究了眼睛表型，预期这些小鼠在表面外胚层衍生物中显示Pax 6的组织特异性减少。为了更好地与我们先前对Pax 6 +/−眼表型的研究进行比较，将半合子Le-CreTg/−和杂合子Pax 6 fl/+小鼠与CBA/Ca遗传背景杂交。Le-Cre转基因与CBA/Ca回交7代后，在一些半合子Le-CreTg/−; Pax 6 +/+对照（没有floxed Pax 6 fl等位基因）以及实验Le-CreTg/−; Pax 6 fl/+小鼠中发生了显著的眼睛异常。然而，在Le-Cre-/-; Pax 6 fl/+或Le-Cre-/-; Pax 6 +/+对照（没有Le-Cre转基因）中没有观察到异常。Le-CreTg/−; Pax 6 +/+对照小鼠中眼睛异常的严重程度和频率在Le-CreTg/−小鼠与原始FVB/N品系回交两代后降低，表明该效应是可逆的。这种遗传背景效应表明，眼睛异常是Le-Cre转基因与某些遗传背景中存在的未知修饰基因的等位基因之间相互作用的结果。通过在CBA/Ca背景下引入额外的Pax 6基因拷贝，这些异常也得到了改善，这表明Le-CreTg/−; Pax 6 +/+小鼠中涉及Pax 6耗竭，而不是Cre重组酶对隐蔽的假loxP位点的直接作用。一种可能性是来自Le-Cre转基因中Pax 6-Le调节序列的Cre重组酶的表达耗尽了内源性Pax 6基因表达所需的辅因子。我们观察到，在没有floxed等位基因的情况下，半合子Le-CreTg/−; Pax 6 +/+小鼠可能发生眼睛异常，这表明在Cre-loxP实验中包括所有相关遗传控制的重要性。
Eye phenotypes were investigated in Le-CreTg/−; Pax6fl/+ mice, which were expected to show tissue-specific reduction of Pax6 in surface ectoderm derivatives. To provide a better comparison with our previous studies of Pax6+/− eye phenotypes, hemizygous Le-CreTg/− and heterozygous Pax6fl/+mice were crossed onto the CBA/Ca genetic background. After the Le-Cre transgene had been backcrossed to CBA/Ca for seven generations, significant eye abnormalities occurred in some hemizygous Le-CreTg/−; Pax6+/+ controls (without a floxed Pax6fl allele) as well as experimental Le-CreTg/−; Pax6fl/+ mice. However, no abnormalities were seen in Le-Cre−/−; Pax6fl/+ or Le-Cre−/−; Pax6+/+ controls (without the Le-Cre transgene). The severity and frequency of the eye abnormalities in Le-CreTg/−; Pax6+/+ control mice diminished after backcrossing Le-CreTg/− mice to the original FVB/N strain for two generations, showing that the effect was reversible. This genetic background effect suggests that the eye abnormalities are a consequence of an interaction between the Le-Cre transgene and alleles of unknown modifier genes present in certain genetic backgrounds. The abnormalities were also ameliorated by introducing additional Pax6 gene copies on a CBA/Ca background, suggesting involvement of Pax6 depletion in Le-CreTg/−; Pax6+/+ mice rather than direct action of Cre recombinase on cryptic pseudo-loxP sites. One possibility is that expression of Cre recombinase from the Pax6-Le regulatory sequences in the Le-Cre transgene depletes cofactors required for endogenous Pax6 gene expression. Our observation that eye abnormalities can occur in hemizygous Le-CreTg/−; Pax6+/+ mice, in the absence of a floxed allele, demonstrates the importance of including all the relevant genetic controls in Cre-loxP experiments.
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DOI：
10.1073/pnas.161144098
发表时间：
2001-08-14
影响因子：
11.1
作者：
Collinson, JM;Quinn, JC;Hill, RE
通讯作者：
Hill, RE
影响因子：
5.3
作者：
Forni, Paolo E.;Scuoppo, Claudio;Ponzetto, Carola
通讯作者：
Ponzetto, Carola
PAX6剂量对角膜发育，生长和伤口愈合的影响。
影响因子：
2.5
作者：
Dora, Natalie;Ou, Jingxing;Kucerova, Romana;Parisi, Ida;West, John D.;Collinson, J. Martin
通讯作者：
Collinson, J. Martin
影响因子：
10.5
作者：
Ashery-Padan, R;Marquardt, T;Gruss, P
通讯作者：
Gruss, P
影响因子：
6
作者：
Buerger, A;Rozhitskaya, O;Jay, PY
通讯作者：
Jay, PY