Use of a Sendai virus-based vector for effcient transduction of pinniped fbroblasts
Generation of induced pluripotent stem (iPS) cells expanded possibilities of pluripotency and early development studies. Generation of order Carnivora iPS cells from dog (Canis lupus familiaris), snow leopard (Panthera uncia), and American mink (Neovison vison) was previously reported. The aim of th...
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| Format: | Article |
| Language: | English |
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Siberian Branch of the Russian Academy of Sciences, Federal Research Center Institute of Cytology and Genetics, The Vavilov Society of Geneticists and Breeders
2019-01-01
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| Series: | Вавиловский журнал генетики и селекции |
| Subjects: | |
| Online Access: | https://vavilov.elpub.ru/jour/article/view/1804 |
| _version_ | 1826550175834832896 |
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| author | V. R. Beklemisheva A. G. Menzorov |
| author_facet | V. R. Beklemisheva A. G. Menzorov |
| author_sort | V. R. Beklemisheva |
| collection | DOAJ |
| description | Generation of induced pluripotent stem (iPS) cells expanded possibilities of pluripotency and early development studies. Generation of order Carnivora iPS cells from dog (Canis lupus familiaris), snow leopard (Panthera uncia), and American mink (Neovison vison) was previously reported. The aim of the current study was to examine conditions of pinniped fbroblast reprogramming. Pinnipeds are representatives of the suborder Caniformia sharing conservative genomes. There are several ways to deliver reprogramming transcription factors: RNA, proteins, plasmids, viral vectors etc. The most effective delivery systems for mouse and human cells are based on viral vectors. We compared a lentiviral vector which integrates into the genome and a Sendai virusbased vector, CytoTune EmGFP Sendai Fluorescence Reporter. The main advantage of Sendai virusbased vectors is that they do not integrate into the genome. We performed delivery of genetic constructions carrying fluorescent proteins to fbroblasts of seven Pinnipeds: northern fur seal (Callorhinus ursinus), Steller sea lion (Eumetopias jubatus), walrus (Odobenus rosmarus), bearded seal (Erignathus barbatus), Baikal seal (Pusa sibirica), ringed seal (Phoca hispida), and spotted seal (Phoca largha). We also transduced American mink (N. vison), human (Homo sapiens), and mouse (Mus musculus) fbroblasts as a control. We showed that the Sendai virusbased transduction system provides transgene expression onetwo orders of magnitude higher than the lentiviral system at a comparable multiplicity of infection. Also, transgene expression after Sendai virusbased transduction is quite stable and changes only slightly at day four compared to day two. These data allow us to suggest that Sendai virusbased vectors are preferable for generation of Pinniped iPS cells. |
| first_indexed | 2024-03-07T16:06:25Z |
| format | Article |
| id | doaj.art-ca656e46dd9448248ec297ee89f10678 |
| institution | Directory Open Access Journal |
| issn | 2500-3259 |
| language | English |
| last_indexed | 2025-03-14T06:33:10Z |
| publishDate | 2019-01-01 |
| publisher | Siberian Branch of the Russian Academy of Sciences, Federal Research Center Institute of Cytology and Genetics, The Vavilov Society of Geneticists and Breeders |
| record_format | Article |
| series | Вавиловский журнал генетики и селекции |
| spelling | doaj.art-ca656e46dd9448248ec297ee89f106782025-03-05T07:59:24ZengSiberian Branch of the Russian Academy of Sciences, Federal Research Center Institute of Cytology and Genetics, The Vavilov Society of Geneticists and BreedersВавиловский журнал генетики и селекции2500-32592019-01-012281020102510.18699/VJ18.445852Use of a Sendai virus-based vector for effcient transduction of pinniped fbroblastsV. R. Beklemisheva0A. G. Menzorov1Institute of Molecular and Cellular Biology, SB RASInstitute of Cytology and Genetics, SB RAS; Novosibirsk State UniversityGeneration of induced pluripotent stem (iPS) cells expanded possibilities of pluripotency and early development studies. Generation of order Carnivora iPS cells from dog (Canis lupus familiaris), snow leopard (Panthera uncia), and American mink (Neovison vison) was previously reported. The aim of the current study was to examine conditions of pinniped fbroblast reprogramming. Pinnipeds are representatives of the suborder Caniformia sharing conservative genomes. There are several ways to deliver reprogramming transcription factors: RNA, proteins, plasmids, viral vectors etc. The most effective delivery systems for mouse and human cells are based on viral vectors. We compared a lentiviral vector which integrates into the genome and a Sendai virusbased vector, CytoTune EmGFP Sendai Fluorescence Reporter. The main advantage of Sendai virusbased vectors is that they do not integrate into the genome. We performed delivery of genetic constructions carrying fluorescent proteins to fbroblasts of seven Pinnipeds: northern fur seal (Callorhinus ursinus), Steller sea lion (Eumetopias jubatus), walrus (Odobenus rosmarus), bearded seal (Erignathus barbatus), Baikal seal (Pusa sibirica), ringed seal (Phoca hispida), and spotted seal (Phoca largha). We also transduced American mink (N. vison), human (Homo sapiens), and mouse (Mus musculus) fbroblasts as a control. We showed that the Sendai virusbased transduction system provides transgene expression onetwo orders of magnitude higher than the lentiviral system at a comparable multiplicity of infection. Also, transgene expression after Sendai virusbased transduction is quite stable and changes only slightly at day four compared to day two. These data allow us to suggest that Sendai virusbased vectors are preferable for generation of Pinniped iPS cells.https://vavilov.elpub.ru/jour/article/view/1804carnivorasealswalrusreprogrammingips cellssendai viruscytotune emgfp sendai fluorescence reporter |
| spellingShingle | V. R. Beklemisheva A. G. Menzorov Use of a Sendai virus-based vector for effcient transduction of pinniped fbroblasts Вавиловский журнал генетики и селекции carnivora seals walrus reprogramming ips cells sendai virus cytotune emgfp sendai fluorescence reporter |
| title | Use of a Sendai virus-based vector for effcient transduction of pinniped fbroblasts |
| title_full | Use of a Sendai virus-based vector for effcient transduction of pinniped fbroblasts |
| title_fullStr | Use of a Sendai virus-based vector for effcient transduction of pinniped fbroblasts |
| title_full_unstemmed | Use of a Sendai virus-based vector for effcient transduction of pinniped fbroblasts |
| title_short | Use of a Sendai virus-based vector for effcient transduction of pinniped fbroblasts |
| title_sort | use of a sendai virus based vector for effcient transduction of pinniped fbroblasts |
| topic | carnivora seals walrus reprogramming ips cells sendai virus cytotune emgfp sendai fluorescence reporter |
| url | https://vavilov.elpub.ru/jour/article/view/1804 |
| work_keys_str_mv | AT vrbeklemisheva useofasendaivirusbasedvectorforeffcienttransductionofpinnipedfbroblasts AT agmenzorov useofasendaivirusbasedvectorforeffcienttransductionofpinnipedfbroblasts |