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Lentiviral gene transfer into human and murine hematopoietic stem cells: size matters
Contemporary biomedical research increasingly depends on techniques to induce or to inhibit expression of genes in hematopoietic stem cells (HSCs) or other primary cells to assess their roles on cellular processes including differentiation, apoptosis and migration. Surprisingly little information is...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4910193/ https://www.ncbi.nlm.nih.gov/pubmed/27306375 http://dx.doi.org/10.1186/s13104-016-2118-z |
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author | Canté-Barrett, Kirsten Mendes, Rui D. Smits, Willem K. van Helsdingen-van Wijk, Yvette M. Pieters, Rob Meijerink, Jules P. P. |
author_facet | Canté-Barrett, Kirsten Mendes, Rui D. Smits, Willem K. van Helsdingen-van Wijk, Yvette M. Pieters, Rob Meijerink, Jules P. P. |
author_sort | Canté-Barrett, Kirsten |
collection | PubMed |
description | Contemporary biomedical research increasingly depends on techniques to induce or to inhibit expression of genes in hematopoietic stem cells (HSCs) or other primary cells to assess their roles on cellular processes including differentiation, apoptosis and migration. Surprisingly little information is available to optimize lentiviral transduction of HSCs. We have therefore carefully optimized transduction of murine and human HSCs by optimizing vector design, serum-free virus production and virus quantitation. We conclude that the viral RNA length, even in relatively small vectors, is an important factor affecting the lentiviral gene transfer on the level of both the virus production and the cellular transduction efficiency. Efficient transfer of large gene sequences into difficult-to-transduce primary cells will benefit from reducing the lentiviral construct size. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13104-016-2118-z) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4910193 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-49101932016-06-17 Lentiviral gene transfer into human and murine hematopoietic stem cells: size matters Canté-Barrett, Kirsten Mendes, Rui D. Smits, Willem K. van Helsdingen-van Wijk, Yvette M. Pieters, Rob Meijerink, Jules P. P. BMC Res Notes Technical Note Contemporary biomedical research increasingly depends on techniques to induce or to inhibit expression of genes in hematopoietic stem cells (HSCs) or other primary cells to assess their roles on cellular processes including differentiation, apoptosis and migration. Surprisingly little information is available to optimize lentiviral transduction of HSCs. We have therefore carefully optimized transduction of murine and human HSCs by optimizing vector design, serum-free virus production and virus quantitation. We conclude that the viral RNA length, even in relatively small vectors, is an important factor affecting the lentiviral gene transfer on the level of both the virus production and the cellular transduction efficiency. Efficient transfer of large gene sequences into difficult-to-transduce primary cells will benefit from reducing the lentiviral construct size. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13104-016-2118-z) contains supplementary material, which is available to authorized users. BioMed Central 2016-06-16 /pmc/articles/PMC4910193/ /pubmed/27306375 http://dx.doi.org/10.1186/s13104-016-2118-z Text en © The Author(s) 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Technical Note Canté-Barrett, Kirsten Mendes, Rui D. Smits, Willem K. van Helsdingen-van Wijk, Yvette M. Pieters, Rob Meijerink, Jules P. P. Lentiviral gene transfer into human and murine hematopoietic stem cells: size matters |
title | Lentiviral gene transfer into human and murine hematopoietic stem cells: size matters |
title_full | Lentiviral gene transfer into human and murine hematopoietic stem cells: size matters |
title_fullStr | Lentiviral gene transfer into human and murine hematopoietic stem cells: size matters |
title_full_unstemmed | Lentiviral gene transfer into human and murine hematopoietic stem cells: size matters |
title_short | Lentiviral gene transfer into human and murine hematopoietic stem cells: size matters |
title_sort | lentiviral gene transfer into human and murine hematopoietic stem cells: size matters |
topic | Technical Note |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4910193/ https://www.ncbi.nlm.nih.gov/pubmed/27306375 http://dx.doi.org/10.1186/s13104-016-2118-z |
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