TY - JOUR T1 - Highly efficient DNA-free gene disruption in the agricultural pest <em>Ceratitis capitata</em> by CRISPR-Cas9 RNPs JF - bioRxiv DO - 10.1101/127506 SP - 127506 AU - Angela Meccariello AU - Simona Maria Monti AU - Alessandra Romanelli AU - Rita Colonna AU - Pasquale Primo AU - Maria Grazia Inghilterra AU - Giuseppe Del Corsano AU - Antonio Ramaglia AU - Giovanni Iazzetti AU - Antonia Chiarore AU - Francesco Patti AU - Svenia D. Heinze AU - Marco Salvemini AU - Helen Lindsay AU - Elena Chiavacci AU - Alexa Burger AU - Mark D. Robinson AU - Christian Mosimann AU - Daniel Bopp AU - Giuseppe Saccone Y1 - 2017/01/01 UR - http://biorxiv.org/content/early/2017/04/18/127506.abstract N2 - The Mediterranean fruitfly Ceratitis capitata (medfly) is an invasive agricultural pest of high economical impact and has become an emerging model for developing new genetic control strategies as alternative to insecticides. Here, we report the successful adaptation of CRISPR-Cas9-based gene disruption in the medfly by injecting in vitro pre-assembled, solubilized Cas9 ribonucleoprotein complexes (RNPs) loaded with gene-specific sgRNAs into early embryos. When targeting the eye pigmentation gene white eye (we), we observed a high rate of somatic mosaicism in surviving G0 adults. Germline transmission of mutated we alleles by G0 animals was on average above 70%, with individual cases achieving a transmission rate of nearly 100%. We further recovered large deletions in the we gene when two sites were simultaneously targeted by two sgRNAs. CRISPR-Cas9 targeting of the Ceratitis ortholog of the Drosophila segmentation paired gene (Ccprd) caused segmental malformations in late embryos and in hatched larvae. Mutant phenotypes correlate with repair by non-homologous end joining (NHEJ) lesions in the two targeted genes. This simple and highly effective Cas9 RNP-based gene editing to introduce mutations in Ceratitis capitata will significantly advance the design and development of new effective strategies for pest control management. ER -