Microarrays of lentiviruses for gene function screens in immortalized and primary cells.
Here we describe lentivirus-infected cell microarrays for the high-throughput screening of gene function in mammalian cells. To create these arrays, we cultured mammalian cells on glass slides 'printed' with lentiviruses pseudotyped as vesicular stomatitis virus glycoprotein, which encode short hairpin RNA or cDNA. Cells that land on the printed 'features' become infected with lentivirus, creating a living array of stably transduced cell clusters within a monolayer of uninfected cells. The small size of the features of the microarrays (300 microm in diameter) allows high-density spotting of lentivirus, permitting thousands of distinct parallel infections on a single glass slide. Because lentiviruses have a wide cellular tropism, including primary cells, lentivirus-infected cell microarrays can be used as a platform for high-throughput screening in a variety of cell types.
Bailey
Steve N
SN
Whitehead Institute for Biomedical Research and Massachusetts Institute of Technology Department of Biology, Nine Cambridge Center, Cambridge, Massachusetts 02142, USA.
Ali
Siraj M
SM
Carpenter
Anne E
AE
Higgins
Caitlin O
CO
Sabatini
David M
DM
D000818:Animals; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D005786:Gene Expression Regulation / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted; D034904:Lamin Type A / Q000235:genetics / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics; D051379:Mice; D046228:Microarray Analysis / Q000379:methods*; D008856:Microscopy, Fluorescence; D010766:Phosphorylation; D011401:Promoter Regions, Genetic / Q000235:genetics; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D019893:Ribosomal Protein S6 Kinases / Q000378:metabolism; D058570:TOR Serine-Threonine Kinases; D018800:Thy-1 Antigens / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D014759:Viral Envelope Proteins / Q000235:genetics; D000097573:Red Fluorescent Protein
D049452:Green Fluorescent Proteins; D034904:Lamin Type A; D008164:Luminescent Proteins; D008562:Membrane Glycoproteins; D011494:Protein Kinases; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; D019893:Ribosomal Protein S6 Kinases; D058570:TOR Serine-Threonine Kinases; D018800:Thy-1 Antigens; D014759:Viral Envelope Proteins; D000097573:Red Fluorescent Protein; C028391:G protein, vesicular stomatitis virus; C546842:MTOR protein, human; C546843:mTOR protein, mouse