FLP recombinase-mediated site-specific recombination in silkworm, Bombyx mori.
A comprehensive understanding of gene function and the production of site-specific genetically modified mutants are two major goals of genetic engineering in the post-genomic era. Although site-specific recombination systems have been powerful tools for genome manipulation of many organisms, they ha...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2012-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3387143?pdf=render |
| _version_ | 1828434596207788032 |
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| author | Ding-Pei Long Ai-Chun Zhao Xue-Jiao Chen Yang Zhang Wei-Jian Lu Qing Guo Alfred M Handler Zhong-Huai Xiang |
| author_facet | Ding-Pei Long Ai-Chun Zhao Xue-Jiao Chen Yang Zhang Wei-Jian Lu Qing Guo Alfred M Handler Zhong-Huai Xiang |
| author_sort | Ding-Pei Long |
| collection | DOAJ |
| description | A comprehensive understanding of gene function and the production of site-specific genetically modified mutants are two major goals of genetic engineering in the post-genomic era. Although site-specific recombination systems have been powerful tools for genome manipulation of many organisms, they have not yet been established for use in the manipulation of the silkworm Bombyx mori genome. In this study, we achieved site-specific excision of a target gene at predefined chromosomal sites in the silkworm using a FLP/FRT site-specific recombination system. We first constructed two stable transgenic target silkworm strains that both contain a single copy of the transgene construct comprising a target gene expression cassette flanked by FRT sites. Using pre-blastoderm microinjection of a FLP recombinase helper expression vector, 32 G3 site-specific recombinant transgenic individuals were isolated from five of 143 broods. The average frequency of FLP recombinase-mediated site-specific excision in the two target strains genome was approximately 3.5%. This study shows that it is feasible to achieve site-specific recombination in silkworms using the FLP/FRT system. We conclude that the FLP/FRT system is a useful tool for genome manipulation in the silkworm. Furthermore, this is the first reported use of the FLP/FRT system for the genetic manipulation of a lepidopteran genome and thus provides a useful reference for the establishment of genome manipulation technologies in other lepidopteran species. |
| first_indexed | 2024-12-10T18:52:21Z |
| format | Article |
| id | doaj.art-3a908ede7c564abe8f2810c243c55b84 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-10T18:52:21Z |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-3a908ede7c564abe8f2810c243c55b842022-12-22T01:37:16ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0176e4015010.1371/journal.pone.0040150FLP recombinase-mediated site-specific recombination in silkworm, Bombyx mori.Ding-Pei LongAi-Chun ZhaoXue-Jiao ChenYang ZhangWei-Jian LuQing GuoAlfred M HandlerZhong-Huai XiangA comprehensive understanding of gene function and the production of site-specific genetically modified mutants are two major goals of genetic engineering in the post-genomic era. Although site-specific recombination systems have been powerful tools for genome manipulation of many organisms, they have not yet been established for use in the manipulation of the silkworm Bombyx mori genome. In this study, we achieved site-specific excision of a target gene at predefined chromosomal sites in the silkworm using a FLP/FRT site-specific recombination system. We first constructed two stable transgenic target silkworm strains that both contain a single copy of the transgene construct comprising a target gene expression cassette flanked by FRT sites. Using pre-blastoderm microinjection of a FLP recombinase helper expression vector, 32 G3 site-specific recombinant transgenic individuals were isolated from five of 143 broods. The average frequency of FLP recombinase-mediated site-specific excision in the two target strains genome was approximately 3.5%. This study shows that it is feasible to achieve site-specific recombination in silkworms using the FLP/FRT system. We conclude that the FLP/FRT system is a useful tool for genome manipulation in the silkworm. Furthermore, this is the first reported use of the FLP/FRT system for the genetic manipulation of a lepidopteran genome and thus provides a useful reference for the establishment of genome manipulation technologies in other lepidopteran species.http://europepmc.org/articles/PMC3387143?pdf=render |
| spellingShingle | Ding-Pei Long Ai-Chun Zhao Xue-Jiao Chen Yang Zhang Wei-Jian Lu Qing Guo Alfred M Handler Zhong-Huai Xiang FLP recombinase-mediated site-specific recombination in silkworm, Bombyx mori. PLoS ONE |
| title | FLP recombinase-mediated site-specific recombination in silkworm, Bombyx mori. |
| title_full | FLP recombinase-mediated site-specific recombination in silkworm, Bombyx mori. |
| title_fullStr | FLP recombinase-mediated site-specific recombination in silkworm, Bombyx mori. |
| title_full_unstemmed | FLP recombinase-mediated site-specific recombination in silkworm, Bombyx mori. |
| title_short | FLP recombinase-mediated site-specific recombination in silkworm, Bombyx mori. |
| title_sort | flp recombinase mediated site specific recombination in silkworm bombyx mori |
| url | http://europepmc.org/articles/PMC3387143?pdf=render |
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