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基于脂质体的转染增强了非模式线虫系统中的 RNAi 和 CRISPR 介导的诱变。

Liposome-based transfection enhances RNAi and CRISPR-mediated mutagenesis in non-model nematode systems.

机构信息

School of Life Sciences, University of Warwick, Coventry, CV4 7AL, UK.

Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America.

出版信息

Sci Rep. 2019 Jan 24;9(1):483. doi: 10.1038/s41598-018-37036-1.

Abstract

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.

摘要

线虫属于动物界最多样化的门之一。然而,除了少数几个物种外,线虫的功能基因组研究往往在可靠性和成功率方面受到限制。在这里,我们报告说,通过将脂质体技术与显微注射相结合,我们成功地在新描述的线虫属 Auanema 中建立了一系列广泛的基因组技术。该方法还允许 Auanema 的 dauer 幼虫发生可遗传的变化,尽管在显微注射时性腺尚未成熟。作为在其他线虫物种中进行潜在功能研究的概念验证,我们还在自由生活的线虫 Pristionchus pacificus 中诱导了 RNAi,并靶向了人类寄生虫 Strongyloides stercoralis。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1175/6345965/1b93f7ae815a/41598_2018_37036_Fig1_HTML.jpg

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