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Mutations in the pqe-1 Gene Enhance Transgene Expression in Caenorhabditis elegans

Although various genetic tools have been developed and used as transgenes, the expression of the transgenes often is hampered by negative regulators. Disrupting such negative regulators of gene expression is potentially a way to overcome the common problem of low expression of transgenes. To find su...

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Autores principales: Yamada, Koji, Tsuchiya, Jun-ichi, Iino, Yuichi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Genetics Society of America 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3385980/
https://www.ncbi.nlm.nih.gov/pubmed/22870397
http://dx.doi.org/10.1534/g3.112.002832
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author Yamada, Koji
Tsuchiya, Jun-ichi
Iino, Yuichi
author_facet Yamada, Koji
Tsuchiya, Jun-ichi
Iino, Yuichi
author_sort Yamada, Koji
collection PubMed
description Although various genetic tools have been developed and used as transgenes, the expression of the transgenes often is hampered by negative regulators. Disrupting such negative regulators of gene expression is potentially a way to overcome the common problem of low expression of transgenes. To find such regulators whose mutations enhance transgene expression in Caenorhabditis elegans, we took advantage of a newly developed reporter transgene, lin-11pAΔ::venus. This transgene induces expression of a fluorescent protein, Venus, in specific neurons including AIZ, where the expression was stochastic. The frequency of reporter expression in AIZ seemed to be correlated with the strength of transgene expression. By using this system, in which a moderate increase of expression was converted to all-or-none expression states, we describe here a forward genetic screen for mutations that enhance the expression of transgenes. Through the screen, we found that mutations in the pqe-1 gene, which encodes a Q/P-rich nuclear protein with an exonuclease domain, increase the chance of reporter expression in AIZ. The fluorescence intensity in RIC, in which all lin-11pAΔ::venus animals show reporter expression, was increased in pqe-1 mutants, suggesting that pqe-1 reduces the expression level of the transgene. Expression of transgenes with other promoters, 3′UTR, or reporter genes was also enhanced by the pqe-1 mutation, suggesting that the effect was not specific to a particular type of transgenes, whereas the effect did not seem to extend to endogenous genes. We propose that pqe-1 mutants can be used to increase the expression of various useful transgenes.
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spelling pubmed-33859802012-08-07 Mutations in the pqe-1 Gene Enhance Transgene Expression in Caenorhabditis elegans Yamada, Koji Tsuchiya, Jun-ichi Iino, Yuichi G3 (Bethesda) Investigations Although various genetic tools have been developed and used as transgenes, the expression of the transgenes often is hampered by negative regulators. Disrupting such negative regulators of gene expression is potentially a way to overcome the common problem of low expression of transgenes. To find such regulators whose mutations enhance transgene expression in Caenorhabditis elegans, we took advantage of a newly developed reporter transgene, lin-11pAΔ::venus. This transgene induces expression of a fluorescent protein, Venus, in specific neurons including AIZ, where the expression was stochastic. The frequency of reporter expression in AIZ seemed to be correlated with the strength of transgene expression. By using this system, in which a moderate increase of expression was converted to all-or-none expression states, we describe here a forward genetic screen for mutations that enhance the expression of transgenes. Through the screen, we found that mutations in the pqe-1 gene, which encodes a Q/P-rich nuclear protein with an exonuclease domain, increase the chance of reporter expression in AIZ. The fluorescence intensity in RIC, in which all lin-11pAΔ::venus animals show reporter expression, was increased in pqe-1 mutants, suggesting that pqe-1 reduces the expression level of the transgene. Expression of transgenes with other promoters, 3′UTR, or reporter genes was also enhanced by the pqe-1 mutation, suggesting that the effect was not specific to a particular type of transgenes, whereas the effect did not seem to extend to endogenous genes. We propose that pqe-1 mutants can be used to increase the expression of various useful transgenes. Genetics Society of America 2012-07-01 /pmc/articles/PMC3385980/ /pubmed/22870397 http://dx.doi.org/10.1534/g3.112.002832 Text en Copyright © 2012 Yamada et al.
spellingShingle Investigations
Yamada, Koji
Tsuchiya, Jun-ichi
Iino, Yuichi
Mutations in the pqe-1 Gene Enhance Transgene Expression in Caenorhabditis elegans
title Mutations in the pqe-1 Gene Enhance Transgene Expression in Caenorhabditis elegans
title_full Mutations in the pqe-1 Gene Enhance Transgene Expression in Caenorhabditis elegans
title_fullStr Mutations in the pqe-1 Gene Enhance Transgene Expression in Caenorhabditis elegans
title_full_unstemmed Mutations in the pqe-1 Gene Enhance Transgene Expression in Caenorhabditis elegans
title_short Mutations in the pqe-1 Gene Enhance Transgene Expression in Caenorhabditis elegans
title_sort mutations in the pqe-1 gene enhance transgene expression in caenorhabditis elegans
topic Investigations
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3385980/
https://www.ncbi.nlm.nih.gov/pubmed/22870397
http://dx.doi.org/10.1534/g3.112.002832
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