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Abstract：
Giardia is a prevalent single-celled microaerophilic intestinal parasite causing diarrheal disease and significantly impacting global health. Double diploid (essentially tetraploid) Giardia trophozoites have presented a formidable challenge to the development of molecular genetic tools to interrogate gene function. High sequence divergence and the high percentage of hypothetical proteins lacking homology to proteins in other eukaryotes have limited our understanding of Giardia protein function, slowing drug target validation and development. For more than 25 years, Giardia A and B assemblages have been readily amenable to transfection with plasmids or linear DNA templates. Here, we highlight the utility and power of genetic approaches developed to assess protein function in Giardia, with particular emphasis on the more recent clustered regularly interspaced palindromic repeats/Cas9-based methods for knockdowns and knockouts. Robust and reliable molecular genetic approaches are fundamental toward the interrogation of Giardia protein function and evaluation of druggable targets. New genetic approaches tailored for the double diploid Giardia are imperative for understanding Giardia\'s unique biology and pathogenesis.
摘要：
贾第虫是一种常见的单细胞微需氧肠道寄生虫，可引起腹泻病，对全球健康产生重大影响。双二倍体（基本上是四倍体）贾第虫滋养体对分子遗传工具的开发提出了艰巨的挑战，以询问基因功能。高度的序列差异以及与其他真核生物中蛋白质缺乏同源性的假设蛋白质的高百分比限制了我们对贾第虫蛋白功能的理解。减缓药物靶点的验证和开发。超过25年,贾第虫A和B组合易于用质粒或线性DNA模板转染。这里,我们强调了用于评估贾第虫中蛋白质功能的遗传方法的实用性和功效，特别强调最近的成簇规则间隔回文重复/基于Cas9的敲除和敲除方法。稳健可靠的分子遗传学方法是询问贾第虫蛋白功能和评估可药用靶标的基础。为双二倍体贾第虫量身定做的新遗传方法对于理解贾第虫独特的生物学和发病机理是必不可少的。
