CRISPR/Cas9 Mediated Editing of Bdtektin1 Gene Induces Sterility in Male Oriental Fruit Fly, Bactrocera dorsalis (Diptera: Tephritidae)

IF 1.5 4区 农林科学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY
Chikmagalur Nagaraja Bhargava, Karuppannasamy Ashok, Sanjay Kumar Pradhan, Sampath Kumar, Maligeppagol Manamohan, Anil Rai, Ramasamy Asokan
{"title":"CRISPR/Cas9 Mediated Editing of Bdtektin1 Gene Induces Sterility in Male Oriental Fruit Fly, Bactrocera dorsalis (Diptera: Tephritidae)","authors":"Chikmagalur Nagaraja Bhargava,&nbsp;Karuppannasamy Ashok,&nbsp;Sanjay Kumar Pradhan,&nbsp;Sampath Kumar,&nbsp;Maligeppagol Manamohan,&nbsp;Anil Rai,&nbsp;Ramasamy Asokan","doi":"10.1002/arch.70043","DOIUrl":null,"url":null,"abstract":"<div>\n \n <p>The Oriental fruit fly, <i>Bactrocera dorsalis</i> (<i>B. dorsalis</i>) is a highly invasive, widely distributed notorious pest restricting global fruit trade immensely. There are several approaches to managing this pest, still require newer approaches. In this regard, recently a novel approach called precision-guided sterile insect technique (pgSIT) is gaining momentum in inducing both female sex elimination or sex conversion and male sterility at one go. Developing a species-specific pgSIT system requires validation of targets such as sex determination and spermatogenesis genes. In this regard, <i>B. dorsalis</i> is highly amenable for area-wide pest management and in the present study, we have validated the loss-of-function of the spermatogenesis-related gene, <i>tektin1</i> using the CRISPR/Cas9 ribonucleoprotein (RNP) complex. This gene was cloned from the local isolate of <i>B. dorsalis</i> and two promising single guide RNAs (sgRNAs) were designed and validated through in vitro restriction analysis. Injection of the RNP complex (sgRNA + Cas9 protein) into the G0 embryo resulted in three adult males carrying mutations at the target site. The phenotype of the mutants was determined through crossing studies, namely, △1♂ × WT ♀, △2♂ × WT ♀, △3♂ × WT ♀, and WT ♂ × WT ♀ and that showed hatching rates of 0%, 11.70%, 0%, and 45.12%, respectively. The mutant males had more nonviable sperm as compared to control. This study underscores the pivotal role of the <i>Bdtektin1</i> gene for male fertility and is a promising candidate for further development of pgSIT system for <i>B. dorsalis</i>.</p></div>","PeriodicalId":8281,"journal":{"name":"Archives of Insect Biochemistry and Physiology","volume":"118 3","pages":""},"PeriodicalIF":1.5000,"publicationDate":"2025-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Archives of Insect Biochemistry and Physiology","FirstCategoryId":"97","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/arch.70043","RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0

Abstract

The Oriental fruit fly, Bactrocera dorsalis (B. dorsalis) is a highly invasive, widely distributed notorious pest restricting global fruit trade immensely. There are several approaches to managing this pest, still require newer approaches. In this regard, recently a novel approach called precision-guided sterile insect technique (pgSIT) is gaining momentum in inducing both female sex elimination or sex conversion and male sterility at one go. Developing a species-specific pgSIT system requires validation of targets such as sex determination and spermatogenesis genes. In this regard, B. dorsalis is highly amenable for area-wide pest management and in the present study, we have validated the loss-of-function of the spermatogenesis-related gene, tektin1 using the CRISPR/Cas9 ribonucleoprotein (RNP) complex. This gene was cloned from the local isolate of B. dorsalis and two promising single guide RNAs (sgRNAs) were designed and validated through in vitro restriction analysis. Injection of the RNP complex (sgRNA + Cas9 protein) into the G0 embryo resulted in three adult males carrying mutations at the target site. The phenotype of the mutants was determined through crossing studies, namely, △1♂ × WT ♀, △2♂ × WT ♀, △3♂ × WT ♀, and WT ♂ × WT ♀ and that showed hatching rates of 0%, 11.70%, 0%, and 45.12%, respectively. The mutant males had more nonviable sperm as compared to control. This study underscores the pivotal role of the Bdtektin1 gene for male fertility and is a promising candidate for further development of pgSIT system for B. dorsalis.

求助全文
约1分钟内获得全文 求助全文
来源期刊
CiteScore
4.30
自引率
4.50%
发文量
115
审稿时长
12 months
期刊介绍: Archives of Insect Biochemistry and Physiology is an international journal that publishes articles in English that are of interest to insect biochemists and physiologists. Generally these articles will be in, or related to, one of the following subject areas: Behavior, Bioinformatics, Carbohydrates, Cell Line Development, Cell Signalling, Development, Drug Discovery, Endocrinology, Enzymes, Lipids, Molecular Biology, Neurobiology, Nucleic Acids, Nutrition, Peptides, Pharmacology, Pollinators, Proteins, Toxicology. Archives will publish only original articles. Articles that are confirmatory in nature or deal with analytical methods previously described will not be accepted.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信