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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...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Caltech Library
2022
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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. |
format | Online Article Text |
id | pubmed-9634457 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Caltech Library |
record_format | MEDLINE/PubMed |
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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