Microarrays of small molecules embedded in biodegradable polymers for use in mammalian cell-based screens.
We developed a microarray-based system for screening small molecules in mammalian cells. This system is compatible with image-based screens and requires fewer than 100 cells per compound. Each compound is impregnated in a 200-microm-diameter disc composed of biodegradable poly-(D),(L)-lactide/glycolide copolymer. Cells are seeded on top of these discs, and compounds slowly diffuse out, affecting proximal cells. In contrast with microtiter-based screening, this system does not involve the use of wells or walls between each compound-treated group of cells. We demonstrate detection of the effects of a single compound in a large microarray, that diverse compounds can be released in this format, and that extended release over several days is feasible. We performed a small synthetic lethal screen and identified a compound (macbecin II) that has reduced activity in cells with RNA interference-mediated decrease in the expression of tuberous sclerosis 2. Thus, we have developed a microarray-based screening system for testing the effects of small molecules on mammalian cells by using an imaging-based readout. This method will be useful to those performing small-molecule screens to discover new chemical tools and potential therapeutic agents.
Bailey
Steve N
SN
Whitehead Institute for Biomedical Research and Department of Biology, Massachusetts Institute of Technology, 9 Cambridge Center, Cambridge, MA 02142, USA.
Sabatini
David M
DM
Stockwell
Brent R
BR
D001672:Biocompatible Materials; D002460:Cell Line; D004353:Drug Evaluation, Preclinical / Q000295:instrumentation / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D019344:Lactic Acid; D046228:Microarray Analysis / Q000295:instrumentation / Q000379:methods*; D011100:Polyglycolic Acid; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011108:Polymers; D034741:RNA, Small Interfering / Q000235:genetics; D012371:Robotics; D014162:Transfection
D001672:Biocompatible Materials; D011108:Polymers; D034741:RNA, Small Interfering; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid