High-throughput screening of gene function in stem cells using clonal microarrays

Randolph S. Ashton, Joseph Peltier, Christopher A. Fasano, Analeah O'Neill, Joshua Leonard, Sally Temple, David V. Schaffer*, Ravi S. Kane

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

22 Scopus citations

Abstract

We describe a microarray-based approach for the high-throughput screening of gene function in stem cells and demonstrate the potential of this method by growing and isolating clonal populations of both adult and embryonic neural stem cells. Clonal microarrays are constructed by seeding a population of cells at clonal density on micropatterned surfaces generated using soft lithographic microfabrication techniques. Clones of interest can be isolated after assaying in parallel for various cellular processes and functions, including proliferation, signal transduction, and differentiation. We demonstrate the compatibility of the technique with both gain- and loss-of-function studies using cell populations infected with cDNA libraries or DNA constructs that induce RNA interference. The infection of cells with a library prior to seeding and the compact but isolated growth of clonal cell populations will facilitate the screening of large libraries in a wide variety of mammalian cells, including those that are difficult to transfect by conventional methods.

Original languageEnglish (US)
Pages (from-to)2928-2935
Number of pages8
JournalStem Cells
Volume25
Issue number11
DOIs
StatePublished - Nov 2007

Keywords

  • Akt1
  • Neural progenitor cells
  • Soft lithography
  • Sox2 transcription factor

ASJC Scopus subject areas

  • Molecular Medicine
  • Developmental Biology
  • Cell Biology

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