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In vivo enrichment of busulfan-resistant germ cells for efficient production of transgenic avian models
Most transgenic animals are generated using a genome-modified stem cell system and genome modification directly in embryos. Although this system is well-established in the development of transgenic animals, donor cell-derived transgenic animal production is inefficient in some cases. Especially in a...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8080772/ https://www.ncbi.nlm.nih.gov/pubmed/33911174 http://dx.doi.org/10.1038/s41598-021-88706-6 |
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author | Kim, Young Min Park, Kyung Je Park, Jin Se Jung, Kyung Min Han, Jae Yong |
author_facet | Kim, Young Min Park, Kyung Je Park, Jin Se Jung, Kyung Min Han, Jae Yong |
author_sort | Kim, Young Min |
collection | PubMed |
description | Most transgenic animals are generated using a genome-modified stem cell system and genome modification directly in embryos. Although this system is well-established in the development of transgenic animals, donor cell-derived transgenic animal production is inefficient in some cases. Especially in avian models such as chickens, the efficiency of transgenic animal production through primordial germ cells (PGCs) is highly variable compared with embryonic manipulation of mammalian species. Because germ cell and germline-competent stem cell-mediated systems that contain the transgene are enriched only at the upstream level during cell cultivation, the efficiency of transgenic animal production is unreliable. Therefore, we developed an in vivo selection model to enhance the efficiency of transgenic chicken production using microsomal glutathione-S-transferase II (MGSTII)-overexpressing PGCs that are resistant to the alkylating agent busulfan, which induces germ cell-specific cytotoxicity. Under in vitro conditions, MGSTII-tg PGCs were resistant to 1 μM busulfan, which was highly toxic to wild-type PGCs. In germline chimeric roosters, transgene-expressing germ cells were dominantly colonized in the recipient testes after busulfan exposure compared with non-treated germline chimera. In validation of germline transmission, donor PGC-derived progeny production efficiency was 94.68%, and the transgene production rate of heterozygous transgenic chickens was significantly increased in chickens that received 40 mg/kg busulfan (80.33–95.23%) compared with that of non-treated germline chimeras (51.18%). This system is expected to significantly improve the efficiency of generating transgenic chickens and other animal species by increasing the distribution of donor cells in adult testes. |
format | Online Article Text |
id | pubmed-8080772 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-80807722021-04-30 In vivo enrichment of busulfan-resistant germ cells for efficient production of transgenic avian models Kim, Young Min Park, Kyung Je Park, Jin Se Jung, Kyung Min Han, Jae Yong Sci Rep Article Most transgenic animals are generated using a genome-modified stem cell system and genome modification directly in embryos. Although this system is well-established in the development of transgenic animals, donor cell-derived transgenic animal production is inefficient in some cases. Especially in avian models such as chickens, the efficiency of transgenic animal production through primordial germ cells (PGCs) is highly variable compared with embryonic manipulation of mammalian species. Because germ cell and germline-competent stem cell-mediated systems that contain the transgene are enriched only at the upstream level during cell cultivation, the efficiency of transgenic animal production is unreliable. Therefore, we developed an in vivo selection model to enhance the efficiency of transgenic chicken production using microsomal glutathione-S-transferase II (MGSTII)-overexpressing PGCs that are resistant to the alkylating agent busulfan, which induces germ cell-specific cytotoxicity. Under in vitro conditions, MGSTII-tg PGCs were resistant to 1 μM busulfan, which was highly toxic to wild-type PGCs. In germline chimeric roosters, transgene-expressing germ cells were dominantly colonized in the recipient testes after busulfan exposure compared with non-treated germline chimera. In validation of germline transmission, donor PGC-derived progeny production efficiency was 94.68%, and the transgene production rate of heterozygous transgenic chickens was significantly increased in chickens that received 40 mg/kg busulfan (80.33–95.23%) compared with that of non-treated germline chimeras (51.18%). This system is expected to significantly improve the efficiency of generating transgenic chickens and other animal species by increasing the distribution of donor cells in adult testes. Nature Publishing Group UK 2021-04-28 /pmc/articles/PMC8080772/ /pubmed/33911174 http://dx.doi.org/10.1038/s41598-021-88706-6 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Kim, Young Min Park, Kyung Je Park, Jin Se Jung, Kyung Min Han, Jae Yong In vivo enrichment of busulfan-resistant germ cells for efficient production of transgenic avian models |
title | In vivo enrichment of busulfan-resistant germ cells for efficient production of transgenic avian models |
title_full | In vivo enrichment of busulfan-resistant germ cells for efficient production of transgenic avian models |
title_fullStr | In vivo enrichment of busulfan-resistant germ cells for efficient production of transgenic avian models |
title_full_unstemmed | In vivo enrichment of busulfan-resistant germ cells for efficient production of transgenic avian models |
title_short | In vivo enrichment of busulfan-resistant germ cells for efficient production of transgenic avian models |
title_sort | in vivo enrichment of busulfan-resistant germ cells for efficient production of transgenic avian models |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8080772/ https://www.ncbi.nlm.nih.gov/pubmed/33911174 http://dx.doi.org/10.1038/s41598-021-88706-6 |
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