Nucleofection as an efficient nonviral transfection method for human monocytic cells.
Despite some progress in the field of gene transfer into hard-to-transfect cells, so far an efficient nonviral method for monocytes has not been available. A comparison of plasmid DNA with capped and polyadenylated mRNA for enhanced green fluorescent protein gene delivery into the commonly used monocytic cell lines U937 and THP-1 suggested that limited DNA trafficking may be the underlying cause of poor transfection results. As Nucleofector technology delivers DNA (or mRNA) straight into the nucleus, we obtained nucleofection efficiencies of up to 80% without significant cell toxicity. Moreover, as the DNA quickly reaches the nucleus, nucleofected cells were ready for analysis after only 2-6 h. The technique is suitable not only for monocytes but also for other hard-to-transfect cells.
Martinet
Wim
W
Division of Pharmacology, University of Antwerp, Wilrijk, Belgium. martinet@uia.ua.ac.be
Schrijvers
Dorien M
DM
Kockx
Mark M
MM
D002467:Cell Nucleus / Q000235:genetics*; D018274:Electroporation / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D009000:Monocytes / Q000502:physiology*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection / Q000379:methods*
D008164:Luminescent Proteins; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins