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An efficient negative selection marker for Mos1 -mediated single-copy integration in Caenorhabditis elegans

Mos1 -mediated single-copy integration (MosSCI) in C. elegans relies on the introduction of plasmid constructs into the germ line. Such plasmids form extrachromosomal arrays containing multiple copies of the transgene. Presently, one positive-selection and four negative-selection reporters are used...

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Detalles Bibliográficos
Autores principales: Revathi, Kandan, Subramaniam, Kuppuswamy
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Caltech Library 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9634457/
https://www.ncbi.nlm.nih.gov/pubmed/36338151
http://dx.doi.org/10.17912/micropub.biology.000647
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author Revathi, Kandan
Subramaniam, Kuppuswamy
author_facet Revathi, Kandan
Subramaniam, Kuppuswamy
author_sort Revathi, Kandan
collection PubMed
description Mos1 -mediated single-copy integration (MosSCI) in C. elegans relies on the introduction of plasmid constructs into the germ line. Such plasmids form extrachromosomal arrays containing multiple copies of the transgene. Presently, one positive-selection and four negative-selection reporters are used to identify animals that carry the integrated transgene but not the array. Even with four reporters, the negative selection is inefficient. Here, we show that the expression of the toxic protein PEEL-1 from a transgene containing the endogenous peel-1 introns kills all array-carrying animals, which facilitates efficient selection of animals carrying the integrated transgene.
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spelling pubmed-96344572022-11-05 An efficient negative selection marker for Mos1 -mediated single-copy integration in Caenorhabditis elegans Revathi, Kandan Subramaniam, Kuppuswamy MicroPubl Biol New Methods Mos1 -mediated single-copy integration (MosSCI) in C. elegans relies on the introduction of plasmid constructs into the germ line. Such plasmids form extrachromosomal arrays containing multiple copies of the transgene. Presently, one positive-selection and four negative-selection reporters are used to identify animals that carry the integrated transgene but not the array. Even with four reporters, the negative selection is inefficient. Here, we show that the expression of the toxic protein PEEL-1 from a transgene containing the endogenous peel-1 introns kills all array-carrying animals, which facilitates efficient selection of animals carrying the integrated transgene. Caltech Library 2022-10-20 /pmc/articles/PMC9634457/ /pubmed/36338151 http://dx.doi.org/10.17912/micropub.biology.000647 Text en Copyright: © 2022 by the authors https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle New Methods
Revathi, Kandan
Subramaniam, Kuppuswamy
An efficient negative selection marker for Mos1 -mediated single-copy integration in Caenorhabditis elegans
title An efficient negative selection marker for Mos1 -mediated single-copy integration in Caenorhabditis elegans
title_full An efficient negative selection marker for Mos1 -mediated single-copy integration in Caenorhabditis elegans
title_fullStr An efficient negative selection marker for Mos1 -mediated single-copy integration in Caenorhabditis elegans
title_full_unstemmed An efficient negative selection marker for Mos1 -mediated single-copy integration in Caenorhabditis elegans
title_short An efficient negative selection marker for Mos1 -mediated single-copy integration in Caenorhabditis elegans
title_sort efficient negative selection marker for mos1 -mediated single-copy integration in caenorhabditis elegans
topic New Methods
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9634457/
https://www.ncbi.nlm.nih.gov/pubmed/36338151
http://dx.doi.org/10.17912/micropub.biology.000647
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