用于评估tau基因沉默疗法的新型转基因小鼠模型的构建
Generation of a new transgenic mouse model for assessment of tau gene silencing therapies.
作者信息
Fromholt Susan, Reitano Christian, Brown Hilda, Lewis Jada, Borchelt David R
机构信息
Department of Neuroscience, Center for Translational Research in Neurodegenerative Disease, University of Florida, Gainesville, FL, USA.
McKnight Brain Institute, University of Florida, Gainesville, FL, 32610, USA.
出版信息
Alzheimers Res Ther. 2016 Sep 5;8(1):36. doi: 10.1186/s13195-016-0202-1.
BACKGROUND
Targeting the expression of genes has emerged as a potentially viable therapeutic approach to human disease. In Alzheimer's disease, therapies that silence the expression of tau could be a viable strategy to slow disease progression.
METHODS
We produced a novel strain of transgenic mice that could be used to assess the efficacy of gene knockdown therapies for human tau, in live mice. We designed a tetracycline-regulated transgene construct in which the cDNA for human tau was fused to ubiquitin and to luciferase to create a single fusion polyprotein, termed TUL.
RESULTS
When expressed in brain, the TUL polyprotein was cleaved by ubiquitin-processing enzymes to release the luciferase as an independent protein, separating the half-life of luciferase from the long-lived tau protein. Treatment of bigenic tTA/TUL mice with doxycycline produced rapid declines in luciferase levels visualized by in vivo imaging and ex vivo enzyme measurement.
CONCLUSIONS
This new mouse model can be used as a discovery tool in optimizing gene targeting therapeutics directed to reduce human tau mRNA levels.
背景
靶向基因表达已成为一种治疗人类疾病的潜在可行方法。在阿尔茨海默病中，使tau蛋白表达沉默的疗法可能是减缓疾病进展的一种可行策略。
方法
我们培育了一种新型转基因小鼠品系，可用于在活体小鼠中评估针对人类tau蛋白的基因敲低疗法的疗效。我们设计了一种四环素调控的转基因构建体，其中人类tau蛋白的cDNA与泛素和荧光素酶融合，以创建一种单一的融合多蛋白，称为TUL。
结果
当在大脑中表达时，TUL多蛋白被泛素加工酶切割，释放出荧光素酶作为一种独立的蛋白质，从而将荧光素酶的半衰期与长寿的tau蛋白分开。用强力霉素处理双转基因tTA/TUL小鼠，通过体内成像和体外酶测量可见荧光素酶水平迅速下降。
结论
这种新的小鼠模型可作为一种发现工具，用于优化旨在降低人类tau mRNA水平的基因靶向治疗。
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