一种CRISPR/Cas9工程小鼠,携带早期生长应答-1转录因子的条件敲除等位基因。

IF 4.6 Q2 MATERIALS SCIENCE, BIOMATERIALS
Vineet K. Maurya, Yan Ying, Denise G. Lanza, Jason D. Heaney, John P. Lydon
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引用次数: 0

摘要

早期生长反应1(EGR1)介导在许多生理学和病理生理学中对细胞分裂、分化和凋亡不可或缺的转录程序。小鼠全身EGR1敲除(Egr1KO)促进了我们对体内EGR1功能的理解。为了扩大小鼠以组织和/或细胞类型特异性方式研究EGR1反应的实用性,我们生成了一个小鼠模型,其中小鼠EGR1基因的外显子2通过CRISPR/Cas9工程进行固定。混合的Egr1等位基因(Egr1f/f)被设计为能够在小鼠中时空控制Cre介导的Egr1消融。为了证实Egr1f/f等位基因可以使用Cre驱动器消除,我们将Egr1f/f小鼠与全局Cre驱动器杂交,产生Egr1条件敲除(Egr1d/d)小鼠,在该小鼠中,所有组织中的Egr1表达都被切除。基因和蛋白质分析分别证实了Egr1d/d小鼠中外显子2的缺失和EGR1表达的缺失。此外,Egr1d/d雌性表现出先前报道的Egr1KO小鼠明显的生殖表型。因此,本简短技术报告中描述的研究强调了小鼠Egr1固定等位基因在组织和/或细胞类型特异性水平上进一步解决Egr1功能的潜在效用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
A CRISPR/Cas9-engineered mouse carrying a conditional knockout allele for the early growth response-1 transcription factor

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 (Egr1KO) 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 (Egr1f/f) are designed to enable spatiotemporal control of Cre-mediated EGR1 ablation in the mouse. To confirm that the Egr1f/f alleles can be abrogated using a Cre driver, we crossed the Egr1f/f mouse with a global Cre driver to generate the Egr1 conditional knockout (Egr1d/d) 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 Egr1d/d mouse, respectively. Moreover, the Egr1d/d female exhibits overt reproductive phenotypes previously reported for the Egr1KO 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.

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来源期刊
ACS Applied Bio Materials
ACS Applied Bio Materials Chemistry-Chemistry (all)
CiteScore
9.40
自引率
2.10%
发文量
464
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