Duncan Samuel M, Jones Nathaniel G, Mottram Jeremy C
Wellcome Centre for Molecular Parasitology, Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, G12 8TA, UK.
Centre for Immunology and Infection, Department of Biology, University of York, Wentworth Way, Heslington, York, YO10 5DD, UK.
Mol Biochem Parasitol. 2017 Sep;216:30-38. doi: 10.1016/j.molbiopara.2017.06.005. Epub 2017 Jun 16.
In this review we describe the expanding repertoire of molecular tools with which to study gene function in Leishmania. Specifically we review the tools available for studying functions of essential genes, such as plasmid shuffle and DiCre, as well as the rapidly expanding portfolio of available CRISPR/Cas9 approaches for large scale gene knockout and endogenous tagging. We include detail on approaches that allow the direct manipulation of RNA using RNAi and protein levels via Tet or DiCre induced overexpression and destabilization domain mediated degradation. The utilisation of current methods and the development of more advanced molecular tools will lead to greater understanding of the role of essential genes in the parasite and thereby more robust drug target validation, thereby paving the way for the development of novel therapeutics to treat this important disease.
在本综述中，我们描述了用于研究利什曼原虫基因功能的不断扩展的分子工具库。具体而言，我们回顾了可用于研究必需基因功能的工具，如质粒洗牌和DiCre，以及用于大规模基因敲除和内源性标记的快速扩展的CRISPR/Cas9方法组合。我们详细介绍了通过RNA干扰直接操纵RNA以及通过Tet或DiCre诱导过表达和去稳定化结构域介导的降解来调控蛋白质水平的方法。当前方法的应用和更先进分子工具的开发将使我们对寄生虫中必需基因的作用有更深入的了解，从而更有力地验证药物靶点，为开发治疗这种重要疾病的新型疗法铺平道路。
