文章基本信息

标题：Cas9-mediated gene editing in the black-legged tick, Ixodes scapularis, by embryo injection and ReMOT Control
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作者：Arvind Sharma ; Michael N. Pham ; Jeremiah B. Reyes 等
期刊名称：iScience
印刷版ISSN：2589-0042
出版年度：2022
卷号：25
期号：3
页码：1-19
DOI：10.1016/j.isci.2022.103781
语种：English
出版社：Elsevier
摘要：SummaryDespite their capacity to acquire and pass on an array of debilitating pathogens, research on ticks has lagged behind other arthropod vectors, such as mosquitoes, largely because of challenges in applying available genetic and molecular tools. CRISPR-Cas9 is transforming non-model organism research; however, successful gene editing has not yet been reported in ticks. Technical challenges for injecting tick embryos to attempt gene editing have further slowed research progress. Currently, no embryo injection protocol exists for any chelicerate species, including ticks. Herein, we report a successful embryo injection protocol for the black-legged tick,Ixodes scapularis, and the use of this protocol for genome editing with CRISPR-Cas9. We also demonstrate that the ReMOT Control technique could be successfully used to generate genome mutations outside Insecta. Our results provide innovative tools to the tick research community that are essential for advancing our understanding of the molecular mechanisms governing pathogen transmission by tick vectors and the underlying biology of host-vector-pathogen interactions.Graphical abstractDisplay OmittedHighlights•Embryo injection protocol developed forIxodes scapularis•CRISPR/Cas9-mediated gene editing is feasible in ticks•Tick gene editing is possible with embryo injection and adult injection (ReMOT Control)Biological sciences; Biotechnology; Genetic engineering; Biological sciences research methodologies; Biology experimental methods