TY  - JOUR
A1  - Steinbrunn, Torsten
A1  - Chatterjee, Manik
A1  - Bargou, Ralf C.
A1  - Stühmer, Thorsten
T1  - Efficient Transient Transfection of Human Multiple Myeloma Cells by Electroporation - An Appraisal
JF  - PLoS ONE
N2  - 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.
KW  - cell cultures
KW  - green fluorescent protein
KW  - oligonucleotides
KW  - multiple myeloma
KW  - plasmid construction
KW  - transfection
KW  - small interfering RNAs
KW  - electroporation
Y1  - 2014
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-119616
VL  - 9
IS  - 6
ER  -