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Efficient Transient Transfection of Human Multiple Myeloma Cells by Electroporation – An Appraisal

Cell lines represent the everyday workhorses for in vitro research on multiple myeloma (MM) and are regularly employed in all aspects of molecular and pharmacological investigations. Although loss-of-function studies using RNA interference in MM cell lines depend on successful knockdown, no well-est...

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Autores principales: Steinbrunn, Torsten, Chatterjee, Manik, Bargou, Ralf C., Stühmer, Thorsten
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4047019/
https://www.ncbi.nlm.nih.gov/pubmed/24901949
http://dx.doi.org/10.1371/journal.pone.0097443
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author Steinbrunn, Torsten
Chatterjee, Manik
Bargou, Ralf C.
Stühmer, Thorsten
author_facet Steinbrunn, Torsten
Chatterjee, Manik
Bargou, Ralf C.
Stühmer, Thorsten
author_sort Steinbrunn, Torsten
collection PubMed
description Cell lines represent the everyday workhorses for in vitro research on multiple myeloma (MM) and are regularly employed in all aspects of molecular and pharmacological investigations. Although loss-of-function studies using RNA interference in MM cell lines depend on successful knockdown, no well-established and widely applied protocol for efficient transient transfection has so far emerged. Here, we provide an appraisal of electroporation as a means to introduce either short-hairpin RNA expression vectors or synthesised siRNAs into MM cells. We found that electroporation using siRNAs was much more efficient than previously anticipated on the basis of transfection efficiencies deduced from EGFP-expression off protein expression vectors. Such knowledge can even confidently be exploited in “hard-to-transfect” MM cell lines to generate large numbers of transient knockdown phenotype MM cells. In addition, special attention was given to developing a protocol that provides easy implementation, good reproducibility and manageable experimental costs.
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spelling pubmed-40470192014-06-09 Efficient Transient Transfection of Human Multiple Myeloma Cells by Electroporation – An Appraisal Steinbrunn, Torsten Chatterjee, Manik Bargou, Ralf C. Stühmer, Thorsten PLoS One Research Article Cell lines represent the everyday workhorses for in vitro research on multiple myeloma (MM) and are regularly employed in all aspects of molecular and pharmacological investigations. Although loss-of-function studies using RNA interference in MM cell lines depend on successful knockdown, no well-established and widely applied protocol for efficient transient transfection has so far emerged. Here, we provide an appraisal of electroporation as a means to introduce either short-hairpin RNA expression vectors or synthesised siRNAs into MM cells. We found that electroporation using siRNAs was much more efficient than previously anticipated on the basis of transfection efficiencies deduced from EGFP-expression off protein expression vectors. Such knowledge can even confidently be exploited in “hard-to-transfect” MM cell lines to generate large numbers of transient knockdown phenotype MM cells. In addition, special attention was given to developing a protocol that provides easy implementation, good reproducibility and manageable experimental costs. Public Library of Science 2014-06-05 /pmc/articles/PMC4047019/ /pubmed/24901949 http://dx.doi.org/10.1371/journal.pone.0097443 Text en © 2014 Steinbrunn 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
Steinbrunn, Torsten
Chatterjee, Manik
Bargou, Ralf C.
Stühmer, Thorsten
Efficient Transient Transfection of Human Multiple Myeloma Cells by Electroporation – An Appraisal
title Efficient Transient Transfection of Human Multiple Myeloma Cells by Electroporation – An Appraisal
title_full Efficient Transient Transfection of Human Multiple Myeloma Cells by Electroporation – An Appraisal
title_fullStr Efficient Transient Transfection of Human Multiple Myeloma Cells by Electroporation – An Appraisal
title_full_unstemmed Efficient Transient Transfection of Human Multiple Myeloma Cells by Electroporation – An Appraisal
title_short Efficient Transient Transfection of Human Multiple Myeloma Cells by Electroporation – An Appraisal
title_sort efficient transient transfection of human multiple myeloma cells by electroporation – an appraisal
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4047019/
https://www.ncbi.nlm.nih.gov/pubmed/24901949
http://dx.doi.org/10.1371/journal.pone.0097443
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