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Journal of Biomolecular Screening
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High Throughput Screening Method for Identification of New Lipofection Reagents

Anne E. Regelin

Tumor Biology Center, Department of Clinical Research/Phospholipids, Freiburg, Germany

Erhard Fernholz

Roche Molecular Biochemicals, Penzberg, Germany

Harald F. Krug

Forschungszentrum Karlsruhe, Department of Toxicology, Eggenstein-Leopoldshafen, Germany

Ulrich Massing

Tumor Biology Center, Department of Clinical Research/Phospholipids, Freiburg, Germany

Lipofection, the transfer of genetic material into cells by means of cationic lipids, is of growing interest for in vitro and in vivo approaches. In order to identify ideal lipofection reagents in a HTS, we have developed an automated lipofection method for the transfer of reporter genes into cells and for determination of the lipofection results. The method has specifically been designed and optimized for 96-well microtiter plates and can successfully be carried out by a pipetting robot with accessory equipment. It consists of two separate parts: (1) pretransfection (preparation of liposomes, formation of lipoplexes, and lipoplex transfer to the cells) and (2) posttransfection (determination of the reporter enzyme activity and the protein content of the transfected cells). Individual steps of the lipofection method were specifically optimized—for example, lipoplex formation and incubation time as well as cell lysis, cell cultivating, and the reporter gene assay. The HTS method facilitates characterization of the transfection properties (efficiency and cytotoxicity) of large numbers of (cationic) lipids in various adherent cell types.

Journal of Biomolecular Screening, Vol. 6, No. 4, 245-254 (2001)
DOI: 10.1177/108705710100600406
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Home page
 Nucleic Acids ResHome page
F.-H. Chang, C.-H. Lee, M.-T. Chen, C.-C. Kuo, Y.-L. Chiang, C.-Y. Hang, and S. Roffler
Surfection: a new platform for transfected cell arrays
Nucleic Acids Res., February 18, 2004; 32(3): e33 - e33.
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