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Establishment of Quantitative PCR Assays for Active Long Interspersed Nuclear Element-1 Subfamilies in Mice and Applications to the Analysis of Aging-Associated Retrotransposition
The retrotransposon long interspersed nuclear element-1 (LINE-1) can autonomously increase its copy number within a host genome through the retrotransposition process. LINE-1 is active in the germline and in neural progenitor cells, and its somatic retrotransposition activity has a broad impact on n...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2020
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7525186/ https://www.ncbi.nlm.nih.gov/pubmed/33193604 http://dx.doi.org/10.3389/fgene.2020.519206 |
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author | Kuroki, Ryota Murata, Yui Fuke, Satoshi Nakachi, Yutaka Nakashima, Jun Kujoth, Gregory C. Prolla, Tomas A. Bundo, Miki Kato, Tadafumi Iwamoto, Kazuya |
author_facet | Kuroki, Ryota Murata, Yui Fuke, Satoshi Nakachi, Yutaka Nakashima, Jun Kujoth, Gregory C. Prolla, Tomas A. Bundo, Miki Kato, Tadafumi Iwamoto, Kazuya |
author_sort | Kuroki, Ryota |
collection | PubMed |
description | The retrotransposon long interspersed nuclear element-1 (LINE-1) can autonomously increase its copy number within a host genome through the retrotransposition process. LINE-1 is active in the germline and in neural progenitor cells, and its somatic retrotransposition activity has a broad impact on neural development and susceptibility to neuropsychiatric disorders. The method to quantify the genomic copy number of LINE-1 would be important in unraveling the role of retrotransposition, especially in the brain. However, because of the species-specific evolution of LINE-1 sequences, methods for quantifying the copy number should be independently developed. Here, we developed a quantitative PCR (qPCR) assay to measure the copy number of active LINE-1 subfamilies in mice. Using the assay, we investigated aging-associated alterations of LINE-1 copy number in several brain regions in wild-type mice and Polg(+/D257A) mice as a model for accelerated aging. We found that aged Polg(+/D257A) mice showed higher levels of the type GfII LINE-1 in the basal ganglia than the wild-type mice did, highlighting the importance of assays that focus on an individual active LINE-1 subfamily. |
format | Online Article Text |
id | pubmed-7525186 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-75251862020-11-13 Establishment of Quantitative PCR Assays for Active Long Interspersed Nuclear Element-1 Subfamilies in Mice and Applications to the Analysis of Aging-Associated Retrotransposition Kuroki, Ryota Murata, Yui Fuke, Satoshi Nakachi, Yutaka Nakashima, Jun Kujoth, Gregory C. Prolla, Tomas A. Bundo, Miki Kato, Tadafumi Iwamoto, Kazuya Front Genet Genetics The retrotransposon long interspersed nuclear element-1 (LINE-1) can autonomously increase its copy number within a host genome through the retrotransposition process. LINE-1 is active in the germline and in neural progenitor cells, and its somatic retrotransposition activity has a broad impact on neural development and susceptibility to neuropsychiatric disorders. The method to quantify the genomic copy number of LINE-1 would be important in unraveling the role of retrotransposition, especially in the brain. However, because of the species-specific evolution of LINE-1 sequences, methods for quantifying the copy number should be independently developed. Here, we developed a quantitative PCR (qPCR) assay to measure the copy number of active LINE-1 subfamilies in mice. Using the assay, we investigated aging-associated alterations of LINE-1 copy number in several brain regions in wild-type mice and Polg(+/D257A) mice as a model for accelerated aging. We found that aged Polg(+/D257A) mice showed higher levels of the type GfII LINE-1 in the basal ganglia than the wild-type mice did, highlighting the importance of assays that focus on an individual active LINE-1 subfamily. Frontiers Media S.A. 2020-09-16 /pmc/articles/PMC7525186/ /pubmed/33193604 http://dx.doi.org/10.3389/fgene.2020.519206 Text en Copyright © 2020 Kuroki, Murata, Fuke, Nakachi, Nakashima, Kujoth, Prolla, Bundo, Kato and Iwamoto. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Genetics Kuroki, Ryota Murata, Yui Fuke, Satoshi Nakachi, Yutaka Nakashima, Jun Kujoth, Gregory C. Prolla, Tomas A. Bundo, Miki Kato, Tadafumi Iwamoto, Kazuya Establishment of Quantitative PCR Assays for Active Long Interspersed Nuclear Element-1 Subfamilies in Mice and Applications to the Analysis of Aging-Associated Retrotransposition |
title | Establishment of Quantitative PCR Assays for Active Long Interspersed Nuclear Element-1 Subfamilies in Mice and Applications to the Analysis of Aging-Associated Retrotransposition |
title_full | Establishment of Quantitative PCR Assays for Active Long Interspersed Nuclear Element-1 Subfamilies in Mice and Applications to the Analysis of Aging-Associated Retrotransposition |
title_fullStr | Establishment of Quantitative PCR Assays for Active Long Interspersed Nuclear Element-1 Subfamilies in Mice and Applications to the Analysis of Aging-Associated Retrotransposition |
title_full_unstemmed | Establishment of Quantitative PCR Assays for Active Long Interspersed Nuclear Element-1 Subfamilies in Mice and Applications to the Analysis of Aging-Associated Retrotransposition |
title_short | Establishment of Quantitative PCR Assays for Active Long Interspersed Nuclear Element-1 Subfamilies in Mice and Applications to the Analysis of Aging-Associated Retrotransposition |
title_sort | establishment of quantitative pcr assays for active long interspersed nuclear element-1 subfamilies in mice and applications to the analysis of aging-associated retrotransposition |
topic | Genetics |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7525186/ https://www.ncbi.nlm.nih.gov/pubmed/33193604 http://dx.doi.org/10.3389/fgene.2020.519206 |
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