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An Efficient Vector System to Modify Cells Genetically
The transfer of foreign genes into mammalian cells has been essential for understanding the functions of genes and mechanisms of genetic diseases, for the production of coding proteins and for gene therapy applications. Currently, the identification and selection of cells that have received transfer...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3214020/ https://www.ncbi.nlm.nih.gov/pubmed/22096482 http://dx.doi.org/10.1371/journal.pone.0026380 |
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author | Han, Huamin Liu, Qingjun He, Wen Ong, Kristy Liu, Xiaoli Gao, Bin |
author_facet | Han, Huamin Liu, Qingjun He, Wen Ong, Kristy Liu, Xiaoli Gao, Bin |
author_sort | Han, Huamin |
collection | PubMed |
description | The transfer of foreign genes into mammalian cells has been essential for understanding the functions of genes and mechanisms of genetic diseases, for the production of coding proteins and for gene therapy applications. Currently, the identification and selection of cells that have received transferred genetic material can be accomplished by methods, including drug selection, reporter enzyme detection and GFP imaging. These methods may confer antibiotic resistance, or be disruptive, or require special equipment. In this study, we labeled genetically modified cells with a cell surface biotinylation tag by co-transfecting cells with BirA, a biotin ligase. The modified cells can be quickly isolated for downstream applications using a simple streptavidin bead method. This system can also be used to screen cells expressing two sets of genes from separate vectors. |
format | Online Article Text |
id | pubmed-3214020 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-32140202011-11-17 An Efficient Vector System to Modify Cells Genetically Han, Huamin Liu, Qingjun He, Wen Ong, Kristy Liu, Xiaoli Gao, Bin PLoS One Research Article The transfer of foreign genes into mammalian cells has been essential for understanding the functions of genes and mechanisms of genetic diseases, for the production of coding proteins and for gene therapy applications. Currently, the identification and selection of cells that have received transferred genetic material can be accomplished by methods, including drug selection, reporter enzyme detection and GFP imaging. These methods may confer antibiotic resistance, or be disruptive, or require special equipment. In this study, we labeled genetically modified cells with a cell surface biotinylation tag by co-transfecting cells with BirA, a biotin ligase. The modified cells can be quickly isolated for downstream applications using a simple streptavidin bead method. This system can also be used to screen cells expressing two sets of genes from separate vectors. Public Library of Science 2011-11-11 /pmc/articles/PMC3214020/ /pubmed/22096482 http://dx.doi.org/10.1371/journal.pone.0026380 Text en Han et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Han, Huamin Liu, Qingjun He, Wen Ong, Kristy Liu, Xiaoli Gao, Bin An Efficient Vector System to Modify Cells Genetically |
title | An Efficient Vector System to Modify Cells Genetically |
title_full | An Efficient Vector System to Modify Cells Genetically |
title_fullStr | An Efficient Vector System to Modify Cells Genetically |
title_full_unstemmed | An Efficient Vector System to Modify Cells Genetically |
title_short | An Efficient Vector System to Modify Cells Genetically |
title_sort | efficient vector system to modify cells genetically |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3214020/ https://www.ncbi.nlm.nih.gov/pubmed/22096482 http://dx.doi.org/10.1371/journal.pone.0026380 |
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