Development of a novel conditional knockdown mouse based on YB-1 protein degradation
Development of a novel conditional knockdown mouse based on YB-1 protein degradation
复制标题
基于YB-1蛋白降解的新型条件敲除小鼠的研制
DOI：
10.1111/gtc.12642
复制
发表时间：
2018
期刊：
影响因子：
2.1
通讯作者：
Miyamoto-Sato Etsuko
中科院分区：
文献类型：
--
作者：
Huang Lijuan;Ozawa Masaaki;Miyamoto-Sato Etsuko
为了阐明疾病的致病机制并建立有效的治疗方法，迫切需要能够动态分析的动物疾病模型。基因敲除小鼠模型和条件遗传工程小鼠模型被开发用于分析与疾病有关的基因和蛋白质。然而，这些方法都有缺点，包括胚胎致死，副作用和效率低。为了解决这个问题，我们创建了一种新的转基因小鼠模型，其中YB 1基因与不稳定结构域（DD）融合，命名为YB 1-DD小鼠。YB-1在整个发育过程中广泛表达，并被认为是细胞存活因子。新合成的DD蛋白通过蛋白酶体途径降解，但它们的降解可以用甲氧苄啶（TMP）阻断。在这项研究中，我们建立了一种新的条件性敲除小鼠模型，能够直接靶向蛋白质降解;该模型导致配体TMP在YB 1杂合子小鼠中对靶蛋白YB-1的剂量依赖性调节。由于这种条件性敲低小鼠模型似乎是功能性的，因此它具有作为基于直接蛋白质降解控制的有用疾病模型的潜力。
To clarify the pathogenic mechanism of disease and establish effective therapies, animal disease models that can be dynamically analyzed are urgently required. Knockout mouse models and conditional genetically engineered mouse models were developed to analyze genes and proteins involved in disease. However, these methods have drawbacks, including embryonic lethality, side effects and low efficiency. To address this issue, we created a novel transgenic mouse model in which theYB1gene was fused with a destabilizing domain (DD), named theYB1‐DDmouse. YB‐1 is widely expressed throughout development and has been implicated as a cell survival factor. Newly synthesized DD proteins are degraded through the proteasome pathway, but their degradation can be blocked with trimethoprim (TMP). In this study, we established a novel conditional knockdown mouse model that enables targeting of protein degradation directly; this model resulted in dose‐dependent regulation of the target protein YB‐1 by the ligand TMP inYB1heterozygous mice. Since this conditional knockdown mouse model appears to be functional, it has potential as a useful disease model based on direct protein degradation control.