Open AccessHigh‐Throughput Screening of Gene Function in Stem Cells Using Clonal Microarrays
Author(s) -
Ashton Randolph S.,
Peltier Joseph,
Fasano Christopher A.,
O'Neill Analeah,
Leonard Joshua,
Temple Sally,
Schaffer David V.,
Kane Ravi S.
Publication year - 2007
Publication title -
stem cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.159
H-Index - 229
eISSN - 1549-4918
pISSN - 1066-5099
DOI - 10.1634/stemcells.2007-0468
Subject(s) - biology , dna microarray , stem cell , electroporation , embryonic stem cell , microarray , microbiology and biotechnology , transfection , population , cellular differentiation , computational biology , gene , genetics , gene expression , demography , sociology
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. Disclosure of potential conflicts of interest is found at the end of this article.