Liposome-based transfection enhances RNAi and CRISPR-mediated mutagenesis in non-model nematode systems
Liposome-based transfection enhances RNAi and CRISPR-mediated mutagenesis in non-model nematode systems
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DOI：
10.1038/s41598-018-37036-1
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发表时间：
2019-01-24
影响因子：
4.6
通讯作者：
Pires-daSilva, Andre
中科院分区：
文献类型：
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作者：
Adams, Sally;Pathak, Prachi;Pires-daSilva, Andre
线虫属于最多样的动物门之一。然而，除少数物种外，线虫的功能基因组研究往往在可靠性和成功方面受到限制。在这里，我们报告了通过结合基于脂质体的技术和显微注射，我们能够在新描述的线虫属Auanema中建立一系列广泛的基因组技术。该方法还允许达尔幼虫发生可遗传的变化，尽管在显微注射时性腺尚未成熟。作为对其他线虫潜在功能研究的概念证明，我们还在自由生活的Pristionchus pacphaus中诱导了RNAi，并针对人类寄生虫Strongyloids stercoralis进行了RNAi。
Nematodes belong to one of the most diverse animal phyla. However, functional genomic studies in nematodes, other than in a few species, have often been limited in their reliability and success. Here we report that by combining liposome-based technology with microinjection, we were able to establish a wide range of genomic techniques in the newly described nematode genus Auanema. The method also allowed heritable changes in dauer larvae of Auanema, despite the immaturity of the gonad at the time of the microinjection. As proof of concept for potential functional studies in other nematode species, we also induced RNAi in the free-living nematode Pristionchus pacificus and targeted the human parasite Strongyloides stercoralis.