Details

Autor(en) / Beteiligte

• Meccariello, Angela
• Tsoumani, Konstantina T.
• Gravina, Andrea
• Primo, Pasquale
• Buonanno, Martina
• Mathiopoulos, Kostas D.
• Saccone, Giuseppe

Titel

Targeted somatic mutagenesis through CRISPR/Cas9 ribonucleoprotein complexes in the olive fruit fly, Bactrocera oleae

Ist Teil von

• Archives of insect biochemistry and physiology, 2020-06, Vol.104 (2), p.e21667-n/a

Ort / Verlag

United States: Wiley Subscription Services, Inc

Erscheinungsjahr

2020

Quelle

Wiley Online Library - AutoHoldings Journals

Beschreibungen/Notizen

• 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. Clustered regularly interspaced short palindromic repeats/Cas9‐mediated mutagenesis of the white gene in the olive fruit fly produced somatic mosaicism in the eye pigmentation. Research Highlights First‐time clustered regularly interspaced short palindromic repeats induced somatic mutations in Bactrocera oleae. Improvement of the existing molecular toolkit in this pest. Provides new opportunities towards the development of novel non‐genetically modified genetic tools for its control.