Highly efficient genome editing by homology-directed repair using Cas9 protein in Ceratitis capitata

Insect Biochemistry and Molecular Biology
Roswitha A AumannIrina Häcker

Abstract

The Mediterranean fruit fly Ceratitis capitata is a highly polyphagous and invasive insect pest, causing enormous economic damage in horticultural systems. A successful and environment-friendly control strategy is the sterile insect technique (SIT) that reduces pest populations through infertile matings with mass-released, sterilized insects. However, the SIT is not readily applicable to each pest species. While transgenic approaches hold great promise to improve critical aspects of the SIT to transfer it to new species, they are suspect to strict or even prohibitive legislation regarding the release of genetically modified (GM) organisms. In contrast, specific mutations created via CRISPR-Cas genome editing are not regulated as GM in the US, and might thus allow creating optimal strains for SIT. Here, we describe highly efficient homology-directed repair genome editing in C. capitata by injecting pre-assembled CRISPR-Cas9 ribonucleoprotein complexes using different guide RNAs and a short single-stranded oligodeoxynucleotide donor to convert an enhanced green fluorescent protein in C. capitata into a blue fluorescent protein. Six out of seven fertile and individually backcrossed G0 individuals generated 57-90% knock-in rate wit...Continue Reading

Citations

Dec 18, 2019·Archives of Insect Biochemistry and Physiology·Konstantina T TsoumaniPhilippos Aris Papathanos
Jan 25, 2020·Journal of Women & Aging·Katarzyna Filar-MierzwaPaulina Aleksander-Szymanowicz
Feb 25, 2020·Archives of Insect Biochemistry and Physiology·Maria-Eleni Gregoriou, Kostas D Mathiopoulos
Feb 27, 2020·Archives of Insect Biochemistry and Physiology·Angela MeccarielloGiuseppe Saccone
Sep 9, 2019·Current Opinion in Insect Science·Suzan MansourianOlena Riabinina
Jan 23, 2021·Nature Communications·Christopher M WardMarc F Schetelig
Jul 3, 2021·International Journal of Molecular Sciences·Ying YanMarc F Schetelig

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