A high-throughput, homogeneous microplate assay for agents that kill mammalian tissue culture cells.
Screens for cytostasis/cytoxicity have considerable value for the discovery of therapeutic agents and the investigation of the biology of apoptosis. For instance, genetic screens for proteins, protein fragments, peptides, RNAs, or chemicals that kill tissue culture cells may aid in identifying new cancer therapeutic targets. A microplate assay for cell death is needed to achieve throughputs sufficient to sift through thousands of agents from expression or chemical libraries. The authors describe a homogeneous assay for cell death in tissue culture cells compatible with 96- or 384-well plates. In combination with a previously described system for retroviral packaging and transduction, nearly 6000 expression library clones could be screened per week in a 96-well plate format. The screening system may also prove useful for chemical screens.
Pierce
Michael
M
Deltagen Proteomics, Salt Lake City, UT 84108, USA.
Wang
Chunwei
C
Rebentisch
Matt
M
Endo
Mark
M
Stump
Mark
M
Kamb
Alexander
A
D017209:Apoptosis; D016923:Cell Death; D002460:Cell Line; D002478:Cells, Cultured; D046508:Culture Techniques; D018076:DNA, Complementary / Q000378:metabolism; D004354:Drug Screening Assays, Antitumor; D015723:Gene Library; D017930:Genes, Reporter; D006801:Humans; D008164:Luminescent Proteins; D018715:Microscopy, Video; D019151:Peptide Library; D010957:Plasmids / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics; D013997:Time Factors; D014162:Transfection
D018076:DNA, Complementary; D008164:Luminescent Proteins; D019151:Peptide Library