Premium
Targeted somatic mutagenesis through CRISPR/Cas9 ribonucleoprotein complexes in the olive fruit fly, Bactrocera oleae
Author(s) -
Meccariello Angela,
Tsoumani Konstantina T.,
Gravina Andrea,
Primo Pasquale,
Buonanno Martina,
Mathiopoulos Kostas D.,
Saccone Giuseppe
Publication year - 2020
Publication title -
archives of insect biochemistry and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.576
H-Index - 66
eISSN - 1520-6327
pISSN - 0739-4462
DOI - 10.1002/arch.21667
Subject(s) - biology , crispr , cas9 , mutagenesis , bactrocera , ribonucleoprotein , genome editing , genetics , gene , trans activating crrna , somatic cell , tephritidae , mutation , botany , pest analysis , rna
The olive fruit fly, Bactrocera oleae (Diptera: Tephritidae), is the most destructive insect pest of olive cultivation, causing significant economic and production losses. Here, we present the establishment of the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 methodology for gene disruption in this species. We performed targeted mutagenesis of the autosomal gene white ( Bo‐we ), by injecting into early embryos in vitro preassembled and solubilized Cas9 ribonucleoprotein complexes loaded with two gene‐specific single‐guide RNAs. Gene disruption of Bo‐we led to somatic mosaicism of the adult eye color. Large eye patches or even an entire eye lost the iridescent reddish color, indicating the successful biallelic mutagenesis in somatic cells. Cas9 induced either indels in each of the two simultaneously targeted Bo‐we sites or a large deletion of the intervening region. This study demonstrates the first efficient implementation of the CRISPR/Cas9 technology in the olive fly, providing new opportunities towards the development of novel genetic tools for its control.