Kudyba Heather M, Cobb David W, Florentin Anat, Krakowiak Michelle, Muralidharan Vasant
Department of Cellular Biology, University of Georgia; Center for Tropical and Emerging Global Diseases, University of Georgia.
Department of Cellular Biology, University of Georgia.
J Vis Exp. 2018 Sep 18(139):57747. doi: 10.3791/57747.
Malaria is a significant cause of morbidity and mortality worldwide. This disease, which primarily affects those living in tropical and subtropical regions, is caused by infection with Plasmodium parasites. The development of more effective drugs to combat malaria can be accelerated by improving our understanding of the biology of this complex parasite. Genetic manipulation of these parasites is key to understanding their biology; however, historically the genome of P. falciparum has been difficult to manipulate. Recently, CRISPR/Cas9 genome editing has been utilized in malaria parasites, allowing for easier protein tagging, generation of conditional protein knockdowns, and deletion of genes. CRISPR/Cas9 genome editing has proven to be a powerful tool for advancing the field of malaria research. Here, we describe a CRISPR/Cas9 method for generating glmS-based conditional knockdown mutants in P. falciparum. This method is highly adaptable to other types of genetic manipulations, including protein tagging and gene knockouts.
疟疾是全球发病和死亡的重要原因。这种疾病主要影响生活在热带和亚热带地区的人群，由感染疟原虫引起。通过增进我们对这种复杂寄生虫生物学特性的了解，可以加速开发更有效的抗疟药物。对这些寄生虫进行基因操作是了解其生物学特性的关键；然而，从历史上看，恶性疟原虫的基因组一直难以操作。最近，CRISPR/Cas9基因组编辑已被用于疟原虫，使得蛋白质标记更容易、条件性蛋白质敲低的产生以及基因的缺失成为可能。CRISPR/Cas9基因组编辑已被证明是推进疟疾研究领域的有力工具。在此，我们描述一种用于在恶性疟原虫中产生基于glmS的条件性敲低突变体的CRISPR/Cas9方法。该方法高度适用于其他类型的基因操作，包括蛋白质标记和基因敲除。
