CRISPR-Cas9 targeted disruption of the yellow ortholog in the housefly identifies the brown body locus

Abstract The classic brown body (bwb) mutation in the housefly Musca domestica impairs normal melanization of the adult cuticle. In Drosophila melanogaster, a reminiscent pigmentation defect results from mutations in the yellow gene encoding dopachrome conversion enzyme (DCE). Here, we demonstrate t...

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Autores principales:	Svenia D. Heinze, Tea Kohlbrenner, Domenica Ippolito, Angela Meccariello, Alexa Burger, Christian Mosimann, Giuseppe Saccone, Daniel Bopp
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/efc5dcc9906c4412afaf3bf225ef5ea1
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spelling	oai:doaj.org-article:efc5dcc9906c4412afaf3bf225ef5ea12021-12-02T12:32:41ZCRISPR-Cas9 targeted disruption of the yellow ortholog in the housefly identifies the brown body locus10.1038/s41598-017-04686-62045-2322https://doaj.org/article/efc5dcc9906c4412afaf3bf225ef5ea12017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-04686-6https://doaj.org/toc/2045-2322Abstract The classic brown body (bwb) mutation in the housefly Musca domestica impairs normal melanization of the adult cuticle. In Drosophila melanogaster, a reminiscent pigmentation defect results from mutations in the yellow gene encoding dopachrome conversion enzyme (DCE). Here, we demonstrate that the bwb locus structurally and functionally represents the yellow ortholog of Musca domestica, MdY. In bwb Musca strains, we identified two mutant MdY alleles that contain lesions predicted to result in premature truncation of the MdY open reading frame. We targeted wildtype MdY by CRISPR-Cas9 RNPs and generated new mutant alleles that fail to complement existing MdY alleles, genetically confirming that MdY is the bwb locus. We further found evidence for Cas9-mediated interchromosomal recombination between wildtype and mutant bwb alleles. Our work resolves the molecular identity of the classic bwb mutation in Musca domestica and establishes the feasibility of Cas9-mediated genome editing in the Musca model.Svenia D. HeinzeTea KohlbrennerDomenica IppolitoAngela MeccarielloAlexa BurgerChristian MosimannGiuseppe SacconeDaniel BoppNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-9 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Svenia D. Heinze Tea Kohlbrenner Domenica Ippolito Angela Meccariello Alexa Burger Christian Mosimann Giuseppe Saccone Daniel Bopp CRISPR-Cas9 targeted disruption of the yellow ortholog in the housefly identifies the brown body locus
description	Abstract The classic brown body (bwb) mutation in the housefly Musca domestica impairs normal melanization of the adult cuticle. In Drosophila melanogaster, a reminiscent pigmentation defect results from mutations in the yellow gene encoding dopachrome conversion enzyme (DCE). Here, we demonstrate that the bwb locus structurally and functionally represents the yellow ortholog of Musca domestica, MdY. In bwb Musca strains, we identified two mutant MdY alleles that contain lesions predicted to result in premature truncation of the MdY open reading frame. We targeted wildtype MdY by CRISPR-Cas9 RNPs and generated new mutant alleles that fail to complement existing MdY alleles, genetically confirming that MdY is the bwb locus. We further found evidence for Cas9-mediated interchromosomal recombination between wildtype and mutant bwb alleles. Our work resolves the molecular identity of the classic bwb mutation in Musca domestica and establishes the feasibility of Cas9-mediated genome editing in the Musca model.
format	article
author	Svenia D. Heinze Tea Kohlbrenner Domenica Ippolito Angela Meccariello Alexa Burger Christian Mosimann Giuseppe Saccone Daniel Bopp
author_facet	Svenia D. Heinze Tea Kohlbrenner Domenica Ippolito Angela Meccariello Alexa Burger Christian Mosimann Giuseppe Saccone Daniel Bopp
author_sort	Svenia D. Heinze
title	CRISPR-Cas9 targeted disruption of the yellow ortholog in the housefly identifies the brown body locus
title_short	CRISPR-Cas9 targeted disruption of the yellow ortholog in the housefly identifies the brown body locus
title_full	CRISPR-Cas9 targeted disruption of the yellow ortholog in the housefly identifies the brown body locus
title_fullStr	CRISPR-Cas9 targeted disruption of the yellow ortholog in the housefly identifies the brown body locus
title_full_unstemmed	CRISPR-Cas9 targeted disruption of the yellow ortholog in the housefly identifies the brown body locus
title_sort	crispr-cas9 targeted disruption of the yellow ortholog in the housefly identifies the brown body locus
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/efc5dcc9906c4412afaf3bf225ef5ea1
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CRISPR-Cas9 targeted disruption of the yellow ortholog in the housefly identifies the brown body locus

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