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Efficient somatic and germline genome engineering of Bactrocera dorsalis by the CRISPR/Cas9 system
Author(s) -
Zhao Santao,
Xing Zengzhu,
Liu Zhonggeng,
Liu Yanhui,
Liu Xiangrui,
Chen Zhe,
Li Jiahui,
Yan Rihui
Publication year - 2019
Publication title -
pest management science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.296
H-Index - 125
eISSN - 1526-4998
pISSN - 1526-498X
DOI - 10.1002/ps.5305
Subject(s) - bactrocera dorsalis , crispr , biology , genetics , germline , gene , cas9 , genome editing , pest analysis , tephritidae , botany
BACKGROUND Bactrocera dorsalis (Hendel), a very destructive insect pest of many fruits and vegetables, is widespread in many Asian countries. To facilitate control of this pest, it is essential to investigate its genetics and gene function using targeted gene disruption. RESULTS Here, we describe successful targeted mutagenesis of the white and transformer genes in B. dorsalis through use of the clustered regularly interspaced short palindromic repeats/CRISPR‐associated 9 (CRISPR/Cas9) system. Co‐injection of the white sgRNA and Cas9 mRNA into B. dorsalis embryos caused eye color change, and the white mutations in the germline were heritable. CRISPR‐mediated knockout of the sex determination gene transformer ( tra ) in B. dorsalis resulted in a male‐biased sex ratio and adult flies with abnormal outer and interior reproductive organs. Small indels and substitutions were induced by CRIRPR for both genes. CONCLUSION Our data demonstrate that somatic and germline genome engineering of the pest B. dorsalis can be performed efficiently using the CRISPR/Cas9 system, opening the door to the use of the CRISPR‐mediated method for functional annotations of genes in B. dorsalis and for its population control using, for example, such as gene drive. © 2018 Society of Chemical Industry