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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...

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Autores principales: Schimmer, Aaron D., Singh, Rashim Pal, Seneviratne, Ayesh K., Thomas, Geethu E., MacLean, Neil, Hurren, Rose
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
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).
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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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