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Efficient Transient Transfection of Human Multiple Myeloma Cells by Electroporation - An Appraisal
Please always quote using this URN: urn:nbn:de:bvb:20-opus-119616
- 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. WeCell 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.…
Author: | Torsten Steinbrunn, Manik Chatterjee, Ralf C. Bargou, Thorsten Stühmer |
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URN: | urn:nbn:de:bvb:20-opus-119616 |
Document Type: | Journal article |
Faculties: | Medizinische Fakultät / Medizinische Klinik und Poliklinik I |
Language: | English |
Parent Title (English): | PLoS ONE |
Year of Completion: | 2014 |
Volume: | 9 |
Issue: | 6 |
Pagenumber: | e97443 |
Source: | PLoS ONE 9(6): e97443. doi:10.1371/journal.pone.0097443 |
DOI: | https://doi.org/10.1371/journal.pone.0097443 |
Pubmed Id: | https://pubmed.ncbi.nlm.nih.gov/24901949 |
Dewey Decimal Classification: | 6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 616 Krankheiten |
Tag: | cell cultures; electroporation; green fluorescent protein; multiple myeloma; oligonucleotides; plasmid construction; small interfering RNAs; transfection |
Release Date: | 2015/11/11 |
Licence (German): | CC BY: Creative-Commons-Lizenz: Namensnennung |