Efficient DIPA-CRISPR-mediated knockout of an eye pigment gene in the white-backed planthopper,Sogatella furcifera  

作　　者：Meng-Qi Zhang Lang-Lang Gong Ya-Qin Zhao Yun-Feng Ma Gui-Jun Long Huan Guo Xuan-Zheng Liu J.Joe Hull Youssef Dewer Chao Yang Ning-Ning Zhang Ming He Peng He 

机构地区：[1]National Key Laboratory of Green Pesticides,Key Laboratory of Green Pesticide and Agricultural Bioengineering,Ministry of Education,Center for R&D of Fine Chemicals of Guizhou University,Guiyang,China [2]Pest Management and Biocontrol Research Unit,US Arid Land Agricultural Research Center,USDA Agricultural Research Services,Maricopa,Arizona,USA [3]Phytotoxicity Research Department,Central Agricultural Pesticide Laboratory,Agricultural Research Center,Dokki,Giza,Egypt [4]Guizhou Jifeng Seed Industry Limited Liability Company,Xingyi,Guizhou Province,China [5]Shandong Facility Horticulture Bioengineering Research Center,Weifang University of Science and Technology,Weifang,Shandong Province,China

出　　处：《Insect Science》2024年第4期1015-1025,共11页昆虫科学（英文版）

基　　金：supported by:the National Natural Science Foundation of China(grants 32370527,for PH,and 32260671,for MH);the Scientific Research Foundation of Guizhou University of China(grant 2017-33,for MH);the Program of Talent Cultivation of Guizhou University(grant(2019)05,for PH);the Science and Technology Support of Guizhou province(grant QKH(2017)2956,for MH);the Program of Introducing Talents of Discipline to Universities of China(111 Program,D20023);the Frontiers Science Center for Asymmetric Synthesis and Medicinal Molecules,Department of Education,Guizhou Province(Qianjiaohe KY number(2020)004).

摘　　要：Although CRISPR/Cas9 has been widely used in insect gene editing,the need for the microinjection of preblastoderm embryos can preclude the technique being used in insect species with eggs that are small,have hard shells,and/or are difficult to collect and maintain outside of their normal environment.Such is the case with Sogatella furcifera,the white-backed planthopper(WBPH),a significant pest of Oryza sativa(rice)that oviposits inside rice stems.Egg extraction from the stem runs the risk of mechanical damage and hatching is heavily influenced by the micro-environment of the rice stem.To bypass these issues,we targeted embryos prior to oviposition via direct parental(DIPA)-CRISPR,in which Cas9 and single-guide RNAs(sgRNAs)for the WBPH eye pigment gene tryptophan 2,3-dioxygenase were injected into the hemocoel of adult females.Females at varying numbers of days posteclosion were evaluated to determine at what stage their oocyte might be most capable of taking up the gene-editing components.An evaluation of the offspring indicated that the highest G0 gene-edited efficacy(56.7%)occurred in females injected 2 d posteclosion,and that those mutations were heritably transmitted to the G1 generation.This study demonstrates the potential utility of DIPA-CRISPR for future gene-editing studies in non-model insect species and can facilitate the development of novel pest management applications.

关 键 词：CRISPR/Cas9 DIPA-CRISPR gene editing tryptophan 2 3-dioxygenase WBPH 

分 类 号：S435.11[农业科学—农业昆虫与害虫防治]