CRISPR/Cas9 Targeted Editing of Osiris24 in Drosophlia melanogaster / 中国生物化学与分子生物学报

ABSTRACT

The gene expression of Osiris is coincident with the timing of chitin deposition. Osiris gene may be involved in the developmental regulation of insect cuticle. The objective of this study is to generate the gene-edited flies with Osiris24 by CRISPR/Cas9-mediated editing system to understand the traits of Osiris24 mutant flies and the expression pattern of Osiris24. Two sgRNA targeted sequences were designed according to the sequence of exon 1 of Osiris24 and inserted into pCFD4 vector backbone. A donor vector with Gal4 protein sequence was constructed. Above two plasmids were mixed and injected into nosCas9 fly embryos to generate GO generation. The results showed that 92.8% GO flies have Gal4 protein insert in genome. Homozygous mutants of Osiris24 were lethal at the embryonic stage or first-instar stage, and no visible phenotype was observed in heterozygous mutants. Osiris24 is expressed throughout larval and pupal stages. At the larval stage, Osiris24 is mainly expressed in the integument, foregut and hind-gut, while Osiris24 is expressed in the integument and wings at the pupal stage. These results indicated that Osiris24 plays an important role in the development of Drosophila. This study provides a research model for in-depth exploration of Osiris gene function.