Hakimi Hassan, Asada Masahito, Kawazu Shin-Ichiro
Department of Disease Control, National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido, Obihiro 080-8555, Japan.
Department of Global Cooperation, National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido, Obihiro 080-8555, Japan.
Vet Sci. 2021 Oct 8;8(10):222. doi: 10.3390/vetsci8100222.
Development of in vitro culture and completion of genome sequencing of several parasites promoted the efforts to establish transfection systems for these parasites to dissect the gene functions. It has been more than a decade since the establishment of first transfection for , the causative agent of bovine babesiosis. However, the number of genes that were targeted by genetic tools in parasites is limited. This is partially due to the low efficiencies of these methods. The recent adaptation of CRISPR/Cas9 for genome editing of can accelerate the efforts for dissecting this parasite's genome and extend the knowledge on biological aspects of erythrocytic and tick stages of . Additionally, ribozyme as a conditional knockdown system is available that could be used for the characterization of essential genes. The development of high throughput genetic tools is needed to dissect the function of multigene families, targeting several genes in a specific pathway, and finally genome-wide identification of essential genes to find novel drug targets. In this review, we summarized the current tools that are available for and the genes that are being targeted by these tools. This may draw a perspective for the future development of genetic tools and pave the way for the identification of novel drugs or vaccine targets.
几种寄生虫体外培养技术的发展以及基因组测序的完成，推动了为这些寄生虫建立转染系统以剖析基因功能的努力。自首次对牛巴贝斯虫病的病原体进行转染至今已有十多年。然而，在巴贝斯虫寄生虫中，利用遗传工具靶向的基因数量有限。这部分是由于这些方法的效率较低。最近CRISPR/Cas9被应用于巴贝斯虫的基因组编辑，这可以加速对该寄生虫基因组的剖析工作，并扩展对其红细胞阶段和蜱虫阶段生物学特性的认识。此外，作为一种条件性敲低系统的巴贝斯虫核酶也已可用，可用于鉴定必需基因。需要开发高通量遗传工具来剖析多基因家族的功能，靶向特定途径中的多个基因，并最终在全基因组范围内鉴定必需基因以寻找新的药物靶点。在本综述中，我们总结了目前可用于巴贝斯虫的工具以及这些工具所靶向的基因。这可能为遗传工具的未来发展提供一个视角，并为鉴定新的药物或疫苗靶点铺平道路。
