Protocols for CRISPR/Cas9 Mutagenesis of the Oriental Fruit Fly Bactrocera dorsalis

文献类型： 外文期刊

第一作者： Yuan, Jinxi

作者： Yuan, Jinxi;Zhang, Jie;Zhang, Yan;QiQiGe, WuYun;Yan, Shanchun;Liu, Wei;Wang, Guirong

作者机构： Northeast Forestry Univ, Key Lab Sustainable Forest Ecosyst Management, Minist Educ, Harbin, Peoples R China;Chinese Acad Agr Sci, Agr Genom Inst Shenzhen, Shenzhen Branch,Minist Agr & Rural Affairs, Guangdong Lab Lingnan Modern Agr,Genome Anal Lab, Beijing, Peoples R China

期刊名称：JOVE-JOURNAL OF VISUALIZED EXPERIMENTS （ 2022影响因子：1.2； 五年影响因子：1.6 ）

ISSN： 1940-087X

年卷期： 2022 年 187 期

页码：

收录情况： SCI

摘要： The Oriental fruit fly, Bactrocera dorsalis, is a highly invasive and adaptive pest species that causes damage to citrus and over 150 other fruit crops worldwide. Since adult fruit flies have great flight capacity and females lay their eggs under the skins of fruit, insecticides requiring direct contact with the pest usually perform poorly in the field. With the development of molecular biological tools and high-throughput sequencing technology, many scientists are attempting to develop environmentally friendly pest management strategies. These include RNAi or gene editing-based pesticides that downregulate or silence genes (molecular targets), such as olfactory genes involved in searching behavior, in various insect pests. To adapt these strategies for Oriental fruit fly control, effective methods for functional gene research are needed. Genes with critical functions in the survival and reproduction of B. dorsalis serve as good molecular targets for gene knockdown and/or silencing. The CRISPR/Cas9 system is a reliable technique used for gene editing, especially in insects. This paper presents a systematic method for CRISPR/Cas9 mutagenesis of B. dorsalis, including the design and synthesis of guide RNAs, collecting embryos, embryo injection, insect rearing, and mutant screening. These protocols will serve as a useful guide for generating mutant flies for researchers interested in functional gene studies in B. dorsalis.

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