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Transduction of Primary AML Cells with Lentiviral Vector for In Vitro Study or In Vivo Engraftment
We describe a method to silence genes in primary acute myeloid leukemia cells by transducing them with shRNA in lentiviral vectors. The transduction of primary non-adherent cells is particularly challenging. The protocol will aid in performing such experiments and is particularly helpful to prepare...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7757357/ https://www.ncbi.nlm.nih.gov/pubmed/33377057 http://dx.doi.org/10.1016/j.xpro.2020.100163 |
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author | Schimmer, Aaron D. Singh, Rashim Pal Seneviratne, Ayesh K. Thomas, Geethu E. MacLean, Neil Hurren, Rose |
author_facet | Schimmer, Aaron D. Singh, Rashim Pal Seneviratne, Ayesh K. Thomas, Geethu E. MacLean, Neil Hurren, Rose |
author_sort | Schimmer, Aaron D. |
collection | PubMed |
description | We describe a method to silence genes in primary acute myeloid leukemia cells by transducing them with shRNA in lentiviral vectors. The transduction of primary non-adherent cells is particularly challenging. The protocol will aid in performing such experiments and is particularly helpful to prepare cells for in vivo engraftment studies. Use of a special medium supplemented with cytokines preserves the viability of the leukemic stem cells and their ability to engraft the marrow of immune-deficient mice. For complete details on the use and execution of this protocol, please refer to Singh et al. (2020). |
format | Online Article Text |
id | pubmed-7757357 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-77573572020-12-28 Transduction of Primary AML Cells with Lentiviral Vector for In Vitro Study or In Vivo Engraftment Schimmer, Aaron D. Singh, Rashim Pal Seneviratne, Ayesh K. Thomas, Geethu E. MacLean, Neil Hurren, Rose STAR Protoc Protocol We describe a method to silence genes in primary acute myeloid leukemia cells by transducing them with shRNA in lentiviral vectors. The transduction of primary non-adherent cells is particularly challenging. The protocol will aid in performing such experiments and is particularly helpful to prepare cells for in vivo engraftment studies. Use of a special medium supplemented with cytokines preserves the viability of the leukemic stem cells and their ability to engraft the marrow of immune-deficient mice. For complete details on the use and execution of this protocol, please refer to Singh et al. (2020). Elsevier 2020-11-25 /pmc/articles/PMC7757357/ /pubmed/33377057 http://dx.doi.org/10.1016/j.xpro.2020.100163 Text en © 2020 The Author(s) http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Protocol Schimmer, Aaron D. Singh, Rashim Pal Seneviratne, Ayesh K. Thomas, Geethu E. MacLean, Neil Hurren, Rose Transduction of Primary AML Cells with Lentiviral Vector for In Vitro Study or In Vivo Engraftment |
title | Transduction of Primary AML Cells with Lentiviral Vector for In Vitro Study or In Vivo Engraftment |
title_full | Transduction of Primary AML Cells with Lentiviral Vector for In Vitro Study or In Vivo Engraftment |
title_fullStr | Transduction of Primary AML Cells with Lentiviral Vector for In Vitro Study or In Vivo Engraftment |
title_full_unstemmed | Transduction of Primary AML Cells with Lentiviral Vector for In Vitro Study or In Vivo Engraftment |
title_short | Transduction of Primary AML Cells with Lentiviral Vector for In Vitro Study or In Vivo Engraftment |
title_sort | transduction of primary aml cells with lentiviral vector for in vitro study or in vivo engraftment |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7757357/ https://www.ncbi.nlm.nih.gov/pubmed/33377057 http://dx.doi.org/10.1016/j.xpro.2020.100163 |
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