A cell spot microarray method for production of high density siRNA
transfection microarrays
BMC Genomics, 12 (1):
162 (2011)DOI: 10.1186/1471-2164-12-162
Abstract
BACKGROUND:High-throughput RNAi screening is widely applied in biological
research, but remains expensive, infrastructure-intensive and conversion
of many assays to HTS applications in microplate format is not feasible.RESULTS:Here,
we describe the optimization of a miniaturized cell spot microarray
(CSMA) method, which facilitates utilization of the transfection
microarray technique for disparate RNAi analyses. To promote rapid
adaptation of the method, the concept has been tested with a panel
of 92 adherent cell types, including primary human cells. We demonstrate
the method in the systematic screening of 492 GPCR coding genes for
impact on growth and survival of cultured human prostate cancer cells.CONCLUSIONS:The
CSMA method facilitates reproducible preparation of highly parallel
cell microarrays for large-scale gene knockdown analyses. This will
be critical towards expanding the cell based functional genetic screens
to include more RNAi constructs, allow combinatorial RNAi analyses,
multi-parametric phenotypic readouts or comparative analysis of many
different cell types.