Scientific Reports (Apr 2017)

Generation of heritable germline mutations in the jewel wasp Nasonia vitripennis using CRISPR/Cas9

  • Ming Li,
  • Lauren Yun Cook Au,
  • Deema Douglah,
  • Abigail Chong,
  • Bradley J. White,
  • Patrick M. Ferree,
  • Omar S. Akbari

DOI
https://doi.org/10.1038/s41598-017-00990-3
Journal volume & issue
Vol. 7, no. 1
pp. 1 – 7

Abstract

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Abstract The revolutionary RNA-guided endonuclease CRISPR/Cas9 system has proven to be a powerful tool for gene editing in a plethora of organisms. Here, utilizing this system we developed an efficient protocol for the generation of heritable germline mutations in the parasitoid jewel wasp, Nasonia vitripennis, a rising insect model organism for the study of evolution, development of axis pattern formation, venom production, haplo-diploid sex determination, and host–symbiont interactions. To establish CRISPR-directed gene editing in N. vitripennis, we targeted a conserved eye pigmentation gene cinnabar, generating several independent heritable germline mutations in this gene. Briefly, to generate these mutants, we developed a protocol to efficiently collect N. vitripennis eggs from a parasitized flesh fly pupa, Sarcophaga bullata, inject these eggs with Cas9/guide RNA mixtures, and transfer injected eggs back into the host to continue development. We also describe a flow for screening mutants and establishing stable mutant strains through genetic crosses. Overall, our results demonstrate that the CRISPR/Cas9 system is a powerful tool for genome manipulation in N. vitripennis, with strong potential for expansion to target critical genes, thus allowing for the investigation of several important biological phenomena in this organism.