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An Assay to Detect In Vivo Y Chromosome Loss in Drosophila Wing Disc Cells

Loss of the Y chromosome in Drosophila has no impact on cell viability and therefore allows us to assay the impact of environmental agents and genetic alterations on chromosomal loss. To detect in vivo chromosome loss in cells of the developing Drosophila wing primordia, we first engineered a Y chro...

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Autores principales: Szabad, Janos, Bellen, Hugo J., Venken, Koen J. T.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Genetics Society of America 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3429924/
https://www.ncbi.nlm.nih.gov/pubmed/22973547
http://dx.doi.org/10.1534/g3.112.002899
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author Szabad, Janos
Bellen, Hugo J.
Venken, Koen J. T.
author_facet Szabad, Janos
Bellen, Hugo J.
Venken, Koen J. T.
author_sort Szabad, Janos
collection PubMed
description Loss of the Y chromosome in Drosophila has no impact on cell viability and therefore allows us to assay the impact of environmental agents and genetic alterations on chromosomal loss. To detect in vivo chromosome loss in cells of the developing Drosophila wing primordia, we first engineered a Y chromosome with an attP docking site. By making use of the ΦC31 integrase system, we site-specifically integrated a genomic transgene encompassing the multiple wing hair (mwh) locus into this attP site, leading to a mwh(+)Y chromosome. This chromosome fully rescues the mwh mutant phenotype, an excellent recessive wing cell marker mutation. Loss of this mwh(+)Y chromosome in wing primordial cells then leads to manifestation of the mwh mutant phenotype in mwh-homozygous cells. The forming mwh clones permit us to quantify the effect of agents and genetic alterations by assaying frequency and size of the mwh mosaic spots. To illustrate the use of the mwh(+)Y loss system, the effects of four known mutagens (X-rays, colchicine, ethyl methanesulfonate, and formaldehyde) and two genetic conditions (loss- and gain-of-function lodestar mutant alleles) are documented. The procedure is simple, sensitive, and inexpensive.
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spelling pubmed-34299242012-09-13 An Assay to Detect In Vivo Y Chromosome Loss in Drosophila Wing Disc Cells Szabad, Janos Bellen, Hugo J. Venken, Koen J. T. G3 (Bethesda) Investigations Loss of the Y chromosome in Drosophila has no impact on cell viability and therefore allows us to assay the impact of environmental agents and genetic alterations on chromosomal loss. To detect in vivo chromosome loss in cells of the developing Drosophila wing primordia, we first engineered a Y chromosome with an attP docking site. By making use of the ΦC31 integrase system, we site-specifically integrated a genomic transgene encompassing the multiple wing hair (mwh) locus into this attP site, leading to a mwh(+)Y chromosome. This chromosome fully rescues the mwh mutant phenotype, an excellent recessive wing cell marker mutation. Loss of this mwh(+)Y chromosome in wing primordial cells then leads to manifestation of the mwh mutant phenotype in mwh-homozygous cells. The forming mwh clones permit us to quantify the effect of agents and genetic alterations by assaying frequency and size of the mwh mosaic spots. To illustrate the use of the mwh(+)Y loss system, the effects of four known mutagens (X-rays, colchicine, ethyl methanesulfonate, and formaldehyde) and two genetic conditions (loss- and gain-of-function lodestar mutant alleles) are documented. The procedure is simple, sensitive, and inexpensive. Genetics Society of America 2012-09-01 /pmc/articles/PMC3429924/ /pubmed/22973547 http://dx.doi.org/10.1534/g3.112.002899 Text en Copyright © 2012 Szabad et al. http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution Unported License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Investigations
Szabad, Janos
Bellen, Hugo J.
Venken, Koen J. T.
An Assay to Detect In Vivo Y Chromosome Loss in Drosophila Wing Disc Cells
title An Assay to Detect In Vivo Y Chromosome Loss in Drosophila Wing Disc Cells
title_full An Assay to Detect In Vivo Y Chromosome Loss in Drosophila Wing Disc Cells
title_fullStr An Assay to Detect In Vivo Y Chromosome Loss in Drosophila Wing Disc Cells
title_full_unstemmed An Assay to Detect In Vivo Y Chromosome Loss in Drosophila Wing Disc Cells
title_short An Assay to Detect In Vivo Y Chromosome Loss in Drosophila Wing Disc Cells
title_sort assay to detect in vivo y chromosome loss in drosophila wing disc cells
topic Investigations
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3429924/
https://www.ncbi.nlm.nih.gov/pubmed/22973547
http://dx.doi.org/10.1534/g3.112.002899
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