Cargando…

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

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 b...

Descripción completa

Detalles Bibliográficos
Autores principales: Heinze, Svenia D., Kohlbrenner, Tea, Ippolito, Domenica, Meccariello, Angela, Burger, Alexa, Mosimann, Christian, Saccone, Giuseppe, Bopp, Daniel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5496933/
https://www.ncbi.nlm.nih.gov/pubmed/28676649
http://dx.doi.org/10.1038/s41598-017-04686-6
_version_ 1783248066156953600
author Heinze, Svenia D.
Kohlbrenner, Tea
Ippolito, Domenica
Meccariello, Angela
Burger, Alexa
Mosimann, Christian
Saccone, Giuseppe
Bopp, Daniel
author_facet Heinze, Svenia D.
Kohlbrenner, Tea
Ippolito, Domenica
Meccariello, Angela
Burger, Alexa
Mosimann, Christian
Saccone, Giuseppe
Bopp, Daniel
author_sort Heinze, Svenia D.
collection PubMed
description 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 Online
Article
Text
id pubmed-5496933
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-54969332017-07-10 CRISPR-Cas9 targeted disruption of the yellow ortholog in the housefly identifies the brown body locus Heinze, Svenia D. Kohlbrenner, Tea Ippolito, Domenica Meccariello, Angela Burger, Alexa Mosimann, Christian Saccone, Giuseppe Bopp, Daniel Sci Rep Article 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. Nature Publishing Group UK 2017-07-04 /pmc/articles/PMC5496933/ /pubmed/28676649 http://dx.doi.org/10.1038/s41598-017-04686-6 Text en © The Author(s) 2017 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Heinze, Svenia D.
Kohlbrenner, Tea
Ippolito, Domenica
Meccariello, Angela
Burger, Alexa
Mosimann, Christian
Saccone, Giuseppe
Bopp, Daniel
CRISPR-Cas9 targeted disruption of the yellow ortholog in the housefly identifies the brown body locus
title 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_short 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
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5496933/
https://www.ncbi.nlm.nih.gov/pubmed/28676649
http://dx.doi.org/10.1038/s41598-017-04686-6
work_keys_str_mv AT heinzesveniad crisprcas9targeteddisruptionoftheyelloworthologinthehouseflyidentifiesthebrownbodylocus
AT kohlbrennertea crisprcas9targeteddisruptionoftheyelloworthologinthehouseflyidentifiesthebrownbodylocus
AT ippolitodomenica crisprcas9targeteddisruptionoftheyelloworthologinthehouseflyidentifiesthebrownbodylocus
AT meccarielloangela crisprcas9targeteddisruptionoftheyelloworthologinthehouseflyidentifiesthebrownbodylocus
AT burgeralexa crisprcas9targeteddisruptionoftheyelloworthologinthehouseflyidentifiesthebrownbodylocus
AT mosimannchristian crisprcas9targeteddisruptionoftheyelloworthologinthehouseflyidentifiesthebrownbodylocus
AT sacconegiuseppe crisprcas9targeteddisruptionoftheyelloworthologinthehouseflyidentifiesthebrownbodylocus
AT boppdaniel crisprcas9targeteddisruptionoftheyelloworthologinthehouseflyidentifiesthebrownbodylocus