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一个携带早期生长反应-1 转录因子条件性敲除等位基因的 CRISPR/Cas9 工程化小鼠。

A CRISPR/Cas9-engineered mouse carrying a conditional knockout allele for the early growth response-1 transcription factor.

机构信息

Department of Molecular and Cellular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, Texas, 77030, USA.

Department of Molecular and Human Genetics, Baylor College of Medicine, One Baylor Plaza, Houston, Texas, 77030, USA.

出版信息

Genesis. 2023 Jul;61(3-4):e23515. doi: 10.1002/dvg.23515. Epub 2023 Mar 22.

Abstract

Early growth response 1 (EGR1) mediates transcriptional programs that are indispensable for cell division, differentiation, and apoptosis in numerous physiologies and pathophysiologies. Whole-body EGR1 knockouts in mice (Egr1 ) have advanced our understanding of EGR1 function in an in vivo context. To extend the utility of the mouse to investigate EGR1 responses in a tissue- and/or cell-type-specific manner, we generated a mouse model in which exon 2 of the mouse Egr1 gene is floxed by CRISPR/Cas9 engineering. The floxed Egr1 alleles (Egr1 ) are designed to enable spatiotemporal control of Cre-mediated EGR1 ablation in the mouse. To confirm that the Egr1 alleles can be abrogated using a Cre driver, we crossed the Egr1 mouse with a global Cre driver to generate the Egr1 conditional knockout (Egr1 ) mouse in which EGR1 expression is ablated in all tissues. Genetic and protein analysis confirmed the absence of exon 2 and loss of EGR1 expression in the Egr1 mouse, respectively. Moreover, the Egr1 female exhibits overt reproductive phenotypes previously reported for the Egr1 mouse. Therefore, studies described in this short technical report underscore the potential utility of the murine Egr1 floxed allele to further resolve EGR1 function at a tissue- and/or cell-type-specific level.

摘要

早期生长反应因子 1(EGR1)介导的转录程序对于细胞分裂、分化和凋亡在许多生理和病理生理过程中都是必不可少的。在整体敲除 EGR1 的小鼠(Egr1 -/-)中,我们对 EGR1 在体内环境中的功能有了更深入的了解。为了扩展小鼠模型在组织和/或细胞类型特异性方面研究 EGR1 反应的用途,我们通过 CRISPR/Cas9 工程构建了一个小鼠模型,其中 EGR1 基因的外显子 2 被 floxed。这些 floxed 的 Egr1 等位基因(Egr1 flox/flox )设计用于时空控制 Cre 介导的 EGR1 缺失。为了确认使用 Cre 驱动子可以消除 Egr1 等位基因,我们将 Egr1 小鼠与全局 Cre 驱动子杂交,生成 Egr1 条件性敲除(Egr1 flox/flox ; CreERT2 )小鼠,其中所有组织中的 EGR1 表达均被消除。遗传和蛋白质分析分别证实了 Egr1 小鼠中缺失外显子 2 和 EGR1 表达的缺失。此外,Egr1 雌性表现出先前报道的 Egr1 小鼠的明显生殖表型。因此,本技术报告中描述的研究强调了使用 Egr1 小鼠 floxed 等位基因在组织和/或细胞类型特异性水平上进一步解析 EGR1 功能的潜力。

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