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

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...

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Autores principales:	Ming Li, Lauren Yun Cook Au, Deema Douglah, Abigail Chong, Bradley J. White, Patrick M. Ferree, Omar S. Akbari
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/544b2d45738148b0a760abe49a827970
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spelling	oai:doaj.org-article:544b2d45738148b0a760abe49a8279702021-12-02T16:07:45ZGeneration of heritable germline mutations in the jewel wasp Nasonia vitripennis using CRISPR/Cas910.1038/s41598-017-00990-32045-2322https://doaj.org/article/544b2d45738148b0a760abe49a8279702017-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-00990-3https://doaj.org/toc/2045-2322Abstract 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.Ming LiLauren Yun Cook AuDeema DouglahAbigail ChongBradley J. WhitePatrick M. FerreeOmar S. AkbariNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-7 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ming Li Lauren Yun Cook Au Deema Douglah Abigail Chong Bradley J. White Patrick M. Ferree Omar S. Akbari Generation of heritable germline mutations in the jewel wasp Nasonia vitripennis using CRISPR/Cas9
description	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.
format	article
author	Ming Li Lauren Yun Cook Au Deema Douglah Abigail Chong Bradley J. White Patrick M. Ferree Omar S. Akbari
author_facet	Ming Li Lauren Yun Cook Au Deema Douglah Abigail Chong Bradley J. White Patrick M. Ferree Omar S. Akbari
author_sort	Ming Li
title	Generation of heritable germline mutations in the jewel wasp Nasonia vitripennis using CRISPR/Cas9
title_short	Generation of heritable germline mutations in the jewel wasp Nasonia vitripennis using CRISPR/Cas9
title_full	Generation of heritable germline mutations in the jewel wasp Nasonia vitripennis using CRISPR/Cas9
title_fullStr	Generation of heritable germline mutations in the jewel wasp Nasonia vitripennis using CRISPR/Cas9
title_full_unstemmed	Generation of heritable germline mutations in the jewel wasp Nasonia vitripennis using CRISPR/Cas9
title_sort	generation of heritable germline mutations in the jewel wasp nasonia vitripennis using crispr/cas9
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/544b2d45738148b0a760abe49a827970
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Generation of heritable germline mutations in the jewel wasp Nasonia vitripennis using CRISPR/Cas9

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