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Generation of transgenic mouse line with prostate-specific expression of codon-improved Cre recombinase

BACKGROUND: Genetically engineered mouse models are useful tools to decipher molecular mechanisms of diseases. As for prostates, a rat probasin promoter has been widely used to drive prostate-specific gene expression. To optimize its codon usage to that of mammals, we used codon-improved Cre recombi...

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Autores principales: Kanayama, Mayuko, Nakao, Kazuki, Horie, Shigeo, Aiba, Atsu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Asian Pacific Prostate Society 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6104291/
https://www.ncbi.nlm.nih.gov/pubmed/30140659
http://dx.doi.org/10.1016/j.prnil.2018.04.003
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author Kanayama, Mayuko
Nakao, Kazuki
Horie, Shigeo
Aiba, Atsu
author_facet Kanayama, Mayuko
Nakao, Kazuki
Horie, Shigeo
Aiba, Atsu
author_sort Kanayama, Mayuko
collection PubMed
description BACKGROUND: Genetically engineered mouse models are useful tools to decipher molecular mechanisms of diseases. As for prostates, a rat probasin promoter has been widely used to drive prostate-specific gene expression. To optimize its codon usage to that of mammals, we used codon-improved Cre recombinase (iCre) for prostate-specific Cre-loxP recombination. MATERIALS AND METHODS: We generated transgenic mice that express iCre driven by conventional probasin promoter in a prostate-specific manner (PB-iCre). Linearized PB-iCre transgene deoxyribonucleic acids (DNAs) were microinjected into pronuclei of fertilized mouse embryos. The integration of the transgene was confirmed by Southern blot analysis. A line of transgenic mice expressing a sufficient amount of iCre mRNA in its prostate was selected. To test recombinase activity of PB-iCre in vivo, its offspring was crossbred with Pten(flox/flox) mice in which murine prostate adenocarcinoma is reported to occur upon excision of loxP-flanked regions. RESULTS: Eight founder animals were obtained, all of which showed germ line integration of PB-iCre transgene by Southern blot analysis. Among them, the prostate from only one line (line 58) expressed a sufficient amount of iCre mRNA. This line was crossbred with Pten(flox/flox) mice to generate PB-iCre58/Pten(flox/flox). As a result, 12-week-old PB-iCre58/Pten(flox/flox) mice presented with prostate adenocarcinoma that was histologically similar to human cribriform prostate cancer of Gleason grade 4. CONCLUSIONS: We have successfully established a transgenic mouse line that expresses iCre in a prostate-specific manner.
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spelling pubmed-61042912018-08-23 Generation of transgenic mouse line with prostate-specific expression of codon-improved Cre recombinase Kanayama, Mayuko Nakao, Kazuki Horie, Shigeo Aiba, Atsu Prostate Int Original Article BACKGROUND: Genetically engineered mouse models are useful tools to decipher molecular mechanisms of diseases. As for prostates, a rat probasin promoter has been widely used to drive prostate-specific gene expression. To optimize its codon usage to that of mammals, we used codon-improved Cre recombinase (iCre) for prostate-specific Cre-loxP recombination. MATERIALS AND METHODS: We generated transgenic mice that express iCre driven by conventional probasin promoter in a prostate-specific manner (PB-iCre). Linearized PB-iCre transgene deoxyribonucleic acids (DNAs) were microinjected into pronuclei of fertilized mouse embryos. The integration of the transgene was confirmed by Southern blot analysis. A line of transgenic mice expressing a sufficient amount of iCre mRNA in its prostate was selected. To test recombinase activity of PB-iCre in vivo, its offspring was crossbred with Pten(flox/flox) mice in which murine prostate adenocarcinoma is reported to occur upon excision of loxP-flanked regions. RESULTS: Eight founder animals were obtained, all of which showed germ line integration of PB-iCre transgene by Southern blot analysis. Among them, the prostate from only one line (line 58) expressed a sufficient amount of iCre mRNA. This line was crossbred with Pten(flox/flox) mice to generate PB-iCre58/Pten(flox/flox). As a result, 12-week-old PB-iCre58/Pten(flox/flox) mice presented with prostate adenocarcinoma that was histologically similar to human cribriform prostate cancer of Gleason grade 4. CONCLUSIONS: We have successfully established a transgenic mouse line that expresses iCre in a prostate-specific manner. Asian Pacific Prostate Society 2018-09 2018-04-27 /pmc/articles/PMC6104291/ /pubmed/30140659 http://dx.doi.org/10.1016/j.prnil.2018.04.003 Text en © 2018 Asian Pacific Prostate Society, Published by Elsevier Korea LLC. http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Original Article
Kanayama, Mayuko
Nakao, Kazuki
Horie, Shigeo
Aiba, Atsu
Generation of transgenic mouse line with prostate-specific expression of codon-improved Cre recombinase
title Generation of transgenic mouse line with prostate-specific expression of codon-improved Cre recombinase
title_full Generation of transgenic mouse line with prostate-specific expression of codon-improved Cre recombinase
title_fullStr Generation of transgenic mouse line with prostate-specific expression of codon-improved Cre recombinase
title_full_unstemmed Generation of transgenic mouse line with prostate-specific expression of codon-improved Cre recombinase
title_short Generation of transgenic mouse line with prostate-specific expression of codon-improved Cre recombinase
title_sort generation of transgenic mouse line with prostate-specific expression of codon-improved cre recombinase
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6104291/
https://www.ncbi.nlm.nih.gov/pubmed/30140659
http://dx.doi.org/10.1016/j.prnil.2018.04.003
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