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使用“敲入”方法实现外源基因的Emx1特异性表达。

Emx1-specific expression of foreign genes using "knock-in" approach.

作者信息

Jin X L, Guo H, Mao C, Atkins N, Wang H, Avasthi P P, Tu Y T, Li Y

机构信息

Department of Molecular and Integrative Physiology, Neuroscience Program, Urbana, Illinois 61801, USA.

出版信息

Biochem Biophys Res Commun. 2000 Apr 21;270(3):978-82. doi: 10.1006/bbrc.2000.2532.

Abstract

Emx1 is a mouse homologue of the Drosophila homeobox gene empty spiracles. Its expression is limited to the neurons in developing and adult cerebral cortex and hippocampus. Because of the highly restricted expression pattern of the Emx1 gene, it would be quite desirable to characterize the promoter of the Emx1 for directing foreign gene expression in the transgenic mouse. We report here that we have achieved the Emx1-specific expression in transgenic mice by inserting the lacZ reporter and cre genes directly into the exon 1 of the Emx1 gene using embryonic stem (ES) cell technology. The distribution of the beta-galactosidase activity in the transgenic mice was consistent with the published results obtained using in situ hybridization and immunohistochemistry. Furthermore, we have demonstrated that Cre protein was present in the cerebral cortex of the transgenic mice and was able to mediate loxP-specific recombination in vitro. The creation of this line of cre transgenic mice, and the demonstration that the insertion site located in the exon 1 of the Emx1 gene could render foreign genes a specific expression pattern restricted to the developing and adult cerebral cortex and hippocampus, should be conducive to further studies of the effect of a gene mutation or overexpression upon the development and plasticity of cerebral cortex and hippocampus.

摘要

Emx1是果蝇同源异型盒基因空气门的小鼠同源物。其表达仅限于发育中和成年大脑皮层及海马体中的神经元。由于Emx1基因的表达模式高度受限,因此非常希望对Emx1的启动子进行表征,以便在转基因小鼠中指导外源基因表达。我们在此报告,通过使用胚胎干细胞(ES)技术将lacZ报告基因和cre基因直接插入Emx1基因的外显子1中,我们在转基因小鼠中实现了Emx1特异性表达。转基因小鼠中β-半乳糖苷酶活性的分布与使用原位杂交和免疫组织化学获得的已发表结果一致。此外,我们已经证明Cre蛋白存在于转基因小鼠的大脑皮层中,并且能够在体外介导loxP特异性重组。这种cre转基因小鼠品系的创建,以及证明位于Emx1基因外显子1中的插入位点可使外源基因具有仅限于发育中和成年大脑皮层及海马体中的特异性表达模式,应有助于进一步研究基因突变或过表达对大脑皮层和海马体发育及可塑性的影响。

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