Tools for the genetic manipulation of Herpetomonas muscarum
Tools for the genetic manipulation of Herpetomonas muscarum
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毒蝇疱疹病毒基因操作工具
DOI：
10.1101/2020.01.03.894162
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Sloan M
中科院分区：
文献类型：
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作者：
Sloan M
锥虫寄生虫是重要的人类和动物疾病（如昏睡病和利什曼病）的病原体。大多数锥虫通过昆虫传播到哺乳动物宿主，通常属于双翅目（或真正的苍蝇）。随着针对媒介杀虫剂和杀锥虫药物的耐药性的报道，需要开发新的传播阻断策略。使用吸血载体本身的研究并不广泛，因此，正在开发新的模型系统来解开昆虫与锥虫的相互作用。其中一个例子是模型双翅目果蝇与其天然锥虫毒蝇单胞菌之间的相互作用。我们之前的工作发现，许多转录组变化是在H.果蝇摄入后的毒蝇伞反映了已知的锥虫病致病因素。在这里，我们描述了一套对寄生虫进行基因操纵的工具，从而从这种关联的双方创建一个真正易于处理的昆虫-寄生虫相互作用系统。其中包括转基因荧光标记寄生虫，以跟踪果蝇肠道中的感染动态，以及可用于生成敲入和敲除菌株的质粒迭代。这份简短报告中提出的工具将有助于进一步表征双翅目动物模型中锥虫的建立。
Trypanosomatid parasites are causative agents of important human and animal diseases such as sleeping sickness and leishmaniasis. Most trypanosomatids are transmitted to their mammalian hosts by insects, often belonging to Diptera (or true flies). With resistance to both vector-targeted pesticides and trypanocidal drugs being reported, there is a need for novel transmission blocking strategies to be developed. Studies using the blood-feeding vectors themselves are not broadly accessible, as such, new model systems are being developed to unpick insect-trypanosmatids interactions. One such case is the interactions between the model dipteranDrosophila melanogasterand its natural trypanosomatidHerpetomonas muscarum. Our previous work has found that much of the transcriptomic changes triggered inH. muscarumafter ingestion byDrosophilareflect what is known for disease-causing trypanosomatids. Here we describe a set of tools to genetically manipulate the parasite and therefore create a truly tractable insect-parasite interaction system from both sides of this association. These include transgenic fluorescently tagged parasites to follow infection dynamics in the fly gut as well as iterations of plasmids that can be used for generating knock-in and knock-out strains. The tools presented in this short report will facilitate further characterization of trypanosomatid establishment in a model dipteran.