Identification of Genomic Safe Harbors in the Anhydrobiotic Cell Line, Pv11
Genomic safe harbors (GSHs) provide ideal integration sites for generating transgenic organisms and cells and can be of great benefit in advancing the basic and applied biology of a particular species. Here we report the identification of GSHs in a dry-preservable insect cell line, Pv11, which deriv...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-02-01
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| Series: | Genes |
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| Online Access: | https://www.mdpi.com/2073-4425/13/3/406 |
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| author | Yugo Miyata Shoko Tokumoto Tomohiko Arai Nurislam Shaikhutdinov Ruslan Deviatiiarov Hiroto Fuse Natalia Gogoleva Sofya Garushyants Alexander Cherkasov Alina Ryabova Guzel Gazizova Richard Cornette Elena Shagimardanova Oleg Gusev Takahiro Kikawada |
| author_facet | Yugo Miyata Shoko Tokumoto Tomohiko Arai Nurislam Shaikhutdinov Ruslan Deviatiiarov Hiroto Fuse Natalia Gogoleva Sofya Garushyants Alexander Cherkasov Alina Ryabova Guzel Gazizova Richard Cornette Elena Shagimardanova Oleg Gusev Takahiro Kikawada |
| author_sort | Yugo Miyata |
| collection | DOAJ |
| description | Genomic safe harbors (GSHs) provide ideal integration sites for generating transgenic organisms and cells and can be of great benefit in advancing the basic and applied biology of a particular species. Here we report the identification of GSHs in a dry-preservable insect cell line, Pv11, which derives from the sleeping chironomid, <i>Polypedilum vanderplanki</i>, and similar to the larvae of its progenitor species exhibits extreme desiccation tolerance. To identify GSHs, we carried out genome analysis of transgenic cell lines established by random integration of exogenous genes and found four candidate loci. Targeted knock-in was performed into these sites and the phenotypes of the resulting transgenic cell lines were examined. Precise integration was achieved for three candidate GSHs, and in all three cases integration did not alter the anhydrobiotic ability or the proliferation rate of the cell lines. We therefore suggest these genomic loci represent GSHs in Pv11 cells. Indeed, we successfully constructed a knock-in system and introduced an expression unit into one of these GSHs. We therefore identified several GSHs in Pv11 cells and developed a new technique for producing transgenic Pv11 cells without affecting the phenotype. |
| first_indexed | 2024-03-09T19:48:13Z |
| format | Article |
| id | doaj.art-5241796d9ba342c5ad7363db7684c54d |
| institution | Directory Open Access Journal |
| issn | 2073-4425 |
| language | English |
| last_indexed | 2024-03-09T19:48:13Z |
| publishDate | 2022-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Genes |
| spelling | doaj.art-5241796d9ba342c5ad7363db7684c54d2023-11-24T01:17:44ZengMDPI AGGenes2073-44252022-02-0113340610.3390/genes13030406Identification of Genomic Safe Harbors in the Anhydrobiotic Cell Line, Pv11Yugo Miyata0Shoko Tokumoto1Tomohiko Arai2Nurislam Shaikhutdinov3Ruslan Deviatiiarov4Hiroto Fuse5Natalia Gogoleva6Sofya Garushyants7Alexander Cherkasov8Alina Ryabova9Guzel Gazizova10Richard Cornette11Elena Shagimardanova12Oleg Gusev13Takahiro Kikawada14Division of Biomaterial Sciences, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization (NARO), Tsukuba 305-0851, JapanDivision of Biomaterial Sciences, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization (NARO), Tsukuba 305-0851, JapanDepartment of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Kashiwa 277-8562, JapanCenter of Life Sciences, Skolkovo Institute of Science and Technology, 121205 Moscow, RussiaRegulatory Genomics Research Center, Institute of Fundamental Medicine and Biology, Kazan Federal University, 420012 Kazan, RussiaDepartment of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Kashiwa 277-8562, JapanRegulatory Genomics Research Center, Institute of Fundamental Medicine and Biology, Kazan Federal University, 420012 Kazan, RussiaCenter of Life Sciences, Skolkovo Institute of Science and Technology, 121205 Moscow, RussiaCenter of Life Sciences, Skolkovo Institute of Science and Technology, 121205 Moscow, RussiaRegulatory Genomics Research Center, Institute of Fundamental Medicine and Biology, Kazan Federal University, 420012 Kazan, RussiaRegulatory Genomics Research Center, Institute of Fundamental Medicine and Biology, Kazan Federal University, 420012 Kazan, RussiaDivision of Biomaterial Sciences, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization (NARO), Tsukuba 305-0851, JapanRegulatory Genomics Research Center, Institute of Fundamental Medicine and Biology, Kazan Federal University, 420012 Kazan, RussiaRegulatory Genomics Research Center, Institute of Fundamental Medicine and Biology, Kazan Federal University, 420012 Kazan, RussiaDivision of Biomaterial Sciences, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization (NARO), Tsukuba 305-0851, JapanGenomic safe harbors (GSHs) provide ideal integration sites for generating transgenic organisms and cells and can be of great benefit in advancing the basic and applied biology of a particular species. Here we report the identification of GSHs in a dry-preservable insect cell line, Pv11, which derives from the sleeping chironomid, <i>Polypedilum vanderplanki</i>, and similar to the larvae of its progenitor species exhibits extreme desiccation tolerance. To identify GSHs, we carried out genome analysis of transgenic cell lines established by random integration of exogenous genes and found four candidate loci. Targeted knock-in was performed into these sites and the phenotypes of the resulting transgenic cell lines were examined. Precise integration was achieved for three candidate GSHs, and in all three cases integration did not alter the anhydrobiotic ability or the proliferation rate of the cell lines. We therefore suggest these genomic loci represent GSHs in Pv11 cells. Indeed, we successfully constructed a knock-in system and introduced an expression unit into one of these GSHs. We therefore identified several GSHs in Pv11 cells and developed a new technique for producing transgenic Pv11 cells without affecting the phenotype.https://www.mdpi.com/2073-4425/13/3/406Pv11 cellsgenomic safe harbor sitesanhydrobiosistransgenesiscell engineering |
| spellingShingle | Yugo Miyata Shoko Tokumoto Tomohiko Arai Nurislam Shaikhutdinov Ruslan Deviatiiarov Hiroto Fuse Natalia Gogoleva Sofya Garushyants Alexander Cherkasov Alina Ryabova Guzel Gazizova Richard Cornette Elena Shagimardanova Oleg Gusev Takahiro Kikawada Identification of Genomic Safe Harbors in the Anhydrobiotic Cell Line, Pv11 Genes Pv11 cells genomic safe harbor sites anhydrobiosis transgenesis cell engineering |
| title | Identification of Genomic Safe Harbors in the Anhydrobiotic Cell Line, Pv11 |
| title_full | Identification of Genomic Safe Harbors in the Anhydrobiotic Cell Line, Pv11 |
| title_fullStr | Identification of Genomic Safe Harbors in the Anhydrobiotic Cell Line, Pv11 |
| title_full_unstemmed | Identification of Genomic Safe Harbors in the Anhydrobiotic Cell Line, Pv11 |
| title_short | Identification of Genomic Safe Harbors in the Anhydrobiotic Cell Line, Pv11 |
| title_sort | identification of genomic safe harbors in the anhydrobiotic cell line pv11 |
| topic | Pv11 cells genomic safe harbor sites anhydrobiosis transgenesis cell engineering |
| url | https://www.mdpi.com/2073-4425/13/3/406 |
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