Department of Translational Neurodegeneration, German Center for Neurodegenerative Diseases (DZNE), 81377, Munich, Germany.
Department of Neurology, Technical University of Munich (TUM), 81675, Munich, Germany.
Curr Neurol Neurosci Rep. 2019 Feb 9;19(2):8. doi: 10.1007/s11910-019-0925-z.
We provide an overview about unbiased screens to identify modifiers of alpha-synuclein (αSyn)-induced toxicity, present the models and the libraries that have been used for screening, and describe how hits from primary screens were selected and validated.
Screens can be classified as either genetic or chemical compound modifier screens, but a few screens do not fit this classification. Most screens addressing αSyn-induced toxicity, including genome-wide overexpressing and deletion, were performed in yeast. More recently, newer methods such as CRISPR-Cas9 became available and were used for screening purposes. Paradoxically, given that αSyn-induced toxicity plays a role in neurological diseases, there is a shortage of human cell-based models for screening. Moreover, most screens used mutant or fluorescently tagged forms of αSyn and only very few screens investigated wild-type αSyn. Particularly, no genome-wide αSyn toxicity screen in human dopaminergic neurons has been published so far. Most unbiased screens for modifiers of αSyn toxicity were performed in yeast, and there is a lack of screens performed in human and particularly dopaminergic cells.
我们提供了一个关于非偏见筛选鉴定α-突触核蛋白（αSyn）诱导毒性调节剂的概述，介绍了用于筛选的模型和文库，并描述了如何选择和验证初筛的命中化合物。
筛选可以分为遗传或化学化合物修饰筛选，但也有一些筛选不适合这种分类。大多数针对αSyn 诱导毒性的筛选，包括全基因组过表达和缺失，都是在酵母中进行的。最近，新的方法如 CRISPR-Cas9 也被用于筛选目的。矛盾的是，鉴于 αSyn 诱导毒性在神经疾病中起作用，目前缺乏用于筛选的基于人类细胞的模型。此外，大多数筛选使用突变或荧光标记形式的 αSyn，只有极少数筛选研究了野生型 αSyn。特别是，目前尚未发表全基因组 αSyn 毒性筛选在人类多巴胺能神经元中的应用。大多数针对 αSyn 毒性调节剂的非偏见筛选都是在酵母中进行的，而在人类特别是多巴胺能细胞中进行的筛选则很少。
