Rantala, J.K. orcid.org/0000-0002-4999-5585, Makela, R., Aaltola, A.-R. et al. (5 more authors) (2011) A cell spot microarray method for production of high density siRNA transfection microarrays. BMC Genomics, 12. 162. ISSN 1471-2164
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 largescale 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.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2011 Rantala et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
Keywords: | Prostate Cancer Cell; Array Spot; Prostate Cancer Sample; siRNA Construct; VCaP Cell |
Dates: |
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Institution: | The University of Sheffield |
Academic Units: | The University of Sheffield > Faculty of Medicine, Dentistry and Health (Sheffield) > Department of Oncology (Sheffield) |
Depositing User: | Symplectic Sheffield |
Date Deposited: | 24 Jun 2019 14:33 |
Last Modified: | 25 Jun 2019 18:54 |
Status: | Published |
Publisher: | BMC |
Refereed: | Yes |
Identification Number: | 10.1186/1471-2164-12-162 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:146989 |