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Gfi1-Cre基因敲入小鼠品系:一种用于内耳毛细胞特异性基因缺失的工具。

Gfi1-Cre knock-in mouse line: A tool for inner ear hair cell-specific gene deletion.

作者信息

Yang Hua, Gan Jean, Xie Xiaoling, Deng Min, Feng Liang, Chen Xiaowei, Gao Zhiqiang, Gan Lin

机构信息

Flaum Eye Institute, University of Rochester, New York 14642, USA.

出版信息

Genesis. 2010 Jun;48(6):400-6. doi: 10.1002/dvg.20632.

Abstract

Gfi1 encodes a zinc-finger transcription factor essential for the development and maintenance of haematopoiesis and the inner ear. In mouse inner ear, Gfi1 expression is confined to hair cells during development and in adulthood. To construct a genetic tool for inner ear hair cell-specific gene deletion, we generated a Gfi1-Cre mouse line by knocking-in Cre coding sequences into the Gfi1 locus and inactivating the endogenous Gfi1. The specificity and efficiency of Gfi1-Cre recombinase-mediated recombination in the developing inner ear was revealed through the expression of the conditional R26R-lacZ reporter gene. The onset of lacZ expression in the Gfi1(Cre/+) inner ear was first detected at E13.5 in the vestibule and at E15.5 in the cochlea, coinciding with the generation of hair cells. Throughout inner ear development, lacZ expression was detected only in hair cells. Thus, Gfi1-Cre knock-in mouse line provides a useful tool for gene manipulations specifically in inner ear hair cells.

摘要

Gfi1编码一种锌指转录因子,对造血和内耳的发育及维持至关重要。在小鼠内耳中,Gfi1的表达在发育过程中和成年期都局限于毛细胞。为构建一种用于内耳毛细胞特异性基因缺失的遗传工具,我们通过将Cre编码序列敲入Gfi1基因座并使内源性Gfi1失活,生成了Gfi1-Cre小鼠品系。通过条件性R26R-lacZ报告基因的表达,揭示了Gfi1-Cre重组酶介导的重组在发育中的内耳中的特异性和效率。在Gfi1(Cre/+)内耳中,lacZ表达首先在E13.5时在前庭被检测到,在E15.5时在耳蜗被检测到,这与毛细胞的产生时间一致。在内耳发育过程中,仅在毛细胞中检测到lacZ表达。因此,Gfi1-Cre敲入小鼠品系为专门在内耳毛细胞中进行基因操作提供了一个有用的工具。

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