Microarrays of lentiviruses for gene function screens in immortalized and primary cells

Nat Methods. 2006 Feb;3(2):117-22. doi: 10.1038/nmeth848.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line, Transformed
  • Cell Line, Tumor
  • Cells, Cultured
  • Dendritic Cells / metabolism
  • Fibroblasts / metabolism
  • Gene Expression Regulation / genetics
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism*
  • HeLa Cells
  • Humans
  • Image Processing, Computer-Assisted
  • Lamin Type A / genetics
  • Lamin Type A / metabolism
  • Lentivirus / genetics*
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Membrane Glycoproteins / genetics
  • Mice
  • Microarray Analysis / methods*
  • Microscopy, Fluorescence
  • Phosphorylation
  • Promoter Regions, Genetic / genetics
  • Protein Kinases / genetics
  • Protein Kinases / metabolism
  • RNA Interference
  • RNA, Small Interfering / genetics
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Red Fluorescent Protein
  • Ribosomal Protein S6 Kinases / metabolism
  • TOR Serine-Threonine Kinases
  • Thy-1 Antigens / genetics
  • Thy-1 Antigens / metabolism
  • Transfection
  • Viral Envelope Proteins / genetics

Substances

  • G protein, vesicular stomatitis virus
  • Lamin Type A
  • Luminescent Proteins
  • Membrane Glycoproteins
  • RNA, Small Interfering
  • Recombinant Fusion Proteins
  • Thy-1 Antigens
  • Viral Envelope Proteins
  • Green Fluorescent Proteins
  • Protein Kinases
  • MTOR protein, human
  • mTOR protein, mouse
  • Ribosomal Protein S6 Kinases
  • TOR Serine-Threonine Kinases