A genetic selection method for expression products that induce apoptosis in adherent mammalian cell lines.
The process of apoptosis is carefully controlled in cells, and different cell types display different sensitivities to pro-apoptotic stimuli. The prospect of exploiting such differences for treatment of diseases such as cancer, via novel therapeutic agents, is extremely attractive. Therefore, genetic selections for novel expression products that kill cells may have considerable value. However, such selections are difficult to devise and perform because the selected cells do not grow. We developed a selection scheme designed to enrich for genetic agents that kill cells. The selection is based on detachment of apoptotic cultured mammalian cells from adherent monolayers. We characterized the properties of these detached cells (floating cells), and various aspects of the selection process. This selection method is potentially applicable to many mammalian cell lines.
Sandrock
R
R
Deltagen Proteomics, Inc., 615 Arapeen Dr., Salt Lake City, UT 84108, USA.
Wheatley
W
W
Drees
E
E
Lualhati
M
M
Kamb
A
A
Caponigro
G
G
D000818:Animals; D017304:Annexin A5 / Q000494:pharmacology; D017209:Apoptosis; D051023:BH3 Interacting Domain Death Agonist Protein; D002352:Carrier Proteins / Q000378:metabolism; D002448:Cell Adhesion; D002460:Cell Line; D004396:Coloring Agents / Q000494:pharmacology; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004247:DNA / Q000032:analysis / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D005434:Flow Cytometry; D015723:Gene Library; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012190:Retroviridae / Q000235:genetics; D013997:Time Factors
D017304:Annexin A5; D051023:BH3 Interacting Domain Death Agonist Protein; C494814:BID protein, human; D002352:Carrier Proteins; D004396:Coloring Agents; D017077:Culture Media, Conditioned; D018076:DNA, Complementary; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D004247:DNA