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Highly efficient transgenesis with miniMos in Caenorhabditis briggsae

Caenorhabditis briggsae as a companion species for Caenorhabditis elegans has played an increasingly important role in study of evolution of development and genome and gene regulation. Aided by the isolation of its sister spices, it has recently been established as a model for speciation study. To t...

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Autores principales: Ding, Qiutao, Ren, Xiaoliang, Li, Runsheng, Chan, Luyan, Ho, Vincy W S, Bi, Yu, Xie, Dongying, Zhao, Zhongying
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9713419/
https://www.ncbi.nlm.nih.gov/pubmed/36171682
http://dx.doi.org/10.1093/g3journal/jkac254
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author Ding, Qiutao
Ren, Xiaoliang
Li, Runsheng
Chan, Luyan
Ho, Vincy W S
Bi, Yu
Xie, Dongying
Zhao, Zhongying
author_facet Ding, Qiutao
Ren, Xiaoliang
Li, Runsheng
Chan, Luyan
Ho, Vincy W S
Bi, Yu
Xie, Dongying
Zhao, Zhongying
author_sort Ding, Qiutao
collection PubMed
description Caenorhabditis briggsae as a companion species for Caenorhabditis elegans has played an increasingly important role in study of evolution of development and genome and gene regulation. Aided by the isolation of its sister spices, it has recently been established as a model for speciation study. To take full advantage of the species for comparative study, an effective transgenesis method especially those with single-copy insertion is important for functional comparison. Here, we improved a transposon-based transgenesis methodology that had been originally developed in C. elegans but worked marginally in C. briggsae. By incorporation of a heat shock step, the transgenesis efficiency in C. briggsae with a single-copy insertion is comparable to that in C. elegans. We used the method to generate 54 independent insertions mostly consisting of a mCherry tag over the C. briggsae genome. We demonstrated the use of the tags in identifying interacting loci responsible for hybrid male sterility between C. briggsae and Caenorhabditis nigoni when combined with the GFP tags we generated previously. Finally, we demonstrated that C. briggsae tolerates the C. elegans toxin, PEEL-1, but not SUP-35, making the latter a potential negative selection marker against extrachromosomal array.
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spelling pubmed-97134192022-12-02 Highly efficient transgenesis with miniMos in Caenorhabditis briggsae Ding, Qiutao Ren, Xiaoliang Li, Runsheng Chan, Luyan Ho, Vincy W S Bi, Yu Xie, Dongying Zhao, Zhongying G3 (Bethesda) Investigation Caenorhabditis briggsae as a companion species for Caenorhabditis elegans has played an increasingly important role in study of evolution of development and genome and gene regulation. Aided by the isolation of its sister spices, it has recently been established as a model for speciation study. To take full advantage of the species for comparative study, an effective transgenesis method especially those with single-copy insertion is important for functional comparison. Here, we improved a transposon-based transgenesis methodology that had been originally developed in C. elegans but worked marginally in C. briggsae. By incorporation of a heat shock step, the transgenesis efficiency in C. briggsae with a single-copy insertion is comparable to that in C. elegans. We used the method to generate 54 independent insertions mostly consisting of a mCherry tag over the C. briggsae genome. We demonstrated the use of the tags in identifying interacting loci responsible for hybrid male sterility between C. briggsae and Caenorhabditis nigoni when combined with the GFP tags we generated previously. Finally, we demonstrated that C. briggsae tolerates the C. elegans toxin, PEEL-1, but not SUP-35, making the latter a potential negative selection marker against extrachromosomal array. Oxford University Press 2022-09-28 /pmc/articles/PMC9713419/ /pubmed/36171682 http://dx.doi.org/10.1093/g3journal/jkac254 Text en © The Author(s) 2022. Published by Oxford University Press on behalf of Genetics Society of America. https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Investigation
Ding, Qiutao
Ren, Xiaoliang
Li, Runsheng
Chan, Luyan
Ho, Vincy W S
Bi, Yu
Xie, Dongying
Zhao, Zhongying
Highly efficient transgenesis with miniMos in Caenorhabditis briggsae
title Highly efficient transgenesis with miniMos in Caenorhabditis briggsae
title_full Highly efficient transgenesis with miniMos in Caenorhabditis briggsae
title_fullStr Highly efficient transgenesis with miniMos in Caenorhabditis briggsae
title_full_unstemmed Highly efficient transgenesis with miniMos in Caenorhabditis briggsae
title_short Highly efficient transgenesis with miniMos in Caenorhabditis briggsae
title_sort highly efficient transgenesis with minimos in caenorhabditis briggsae
topic Investigation
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9713419/
https://www.ncbi.nlm.nih.gov/pubmed/36171682
http://dx.doi.org/10.1093/g3journal/jkac254
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