A cell spot microarray method for high-throughput biology: Dissertation

Juha K. Rantala

Research output: ThesisDissertationCollection of Articles

Abstract

High-throughput screening of cellular effects of RNA interference (RNAi) libraries is now being increasingly applied to explore the role of genes in specific cell biological processes and disease states. However, the technology is still limited to specialty laboratories, due to the requirements for robotic infrastructure, access to expensive reagent libraries, expertise in high-throughput screening assay development, standardization, data analysis and applications. In the future, alternative screening platforms will be required to expand functional large-scale experiments to include more RNAi constructs, allow combinatorial loss-of-function analyses (e.g. genegene or gene-drug interaction), gain-of-function screens, multi-parametric phenotypic readouts or comparative analysis of many different cell types. Such comprehensive perturbation of gene networks in cells will require a major increase in the flexibility of the screening platforms, throughput and reduction of costs. As an alternative for the conventional multi-well based high-throughput screening -platforms, here the development of a novel cell spot microarray method for production of high density siRNA reverse transfection arrays is described. The cell spot microarray platform is distinguished from the majority of other transfection cell microarray techniques by the spatially confined array layout that allow highly parallel screening of large-scale RNAi reagent libraries with assays otherwise difficult or not applicable to high-throughput screening. This study depicts the development of the cell spot microarray method along with biological application examples of high-content immunofluorescence and phenotype based cancer cell biological analyses focusing on the regulation of prostate cancer cell growth, maintenance of genomic integrity in breast cancer cells, and functional analysis of integrin protein-protein interactions in situ.
Original languageEnglish
QualificationDoctor Degree
Awarding Institution
  • University of Turku
Supervisors/Advisors
  • Kallioniemi, Olli, Supervisor, External person
  • Ivaska, Johanna, Supervisor, External person
Place of PublicationTurku
Publisher
Print ISBNs978-951-29-4657-0
Electronic ISBNs978-951-29-4658-7
Publication statusPublished - 2011
MoE publication typeG5 Doctoral dissertation (article)

Fingerprint

RNA Interference
Transfection
High-Throughput Screening Assays
Biological Phenomena
Gene Regulatory Networks
Robotics
Drug Interactions
Integrins
Small Interfering RNA
Genes
Libraries
Fluorescent Antibody Technique
Prostatic Neoplasms
Proteins
Maintenance
Breast Neoplasms
Technology
Phenotype
Costs and Cost Analysis
Growth

Keywords

  • Hihg-throughput screening
  • RNA interference
  • cell microarrays
  • RNA interferenssi
  • suurtehoseulonta
  • solumikrosirumenetelmä

Cite this

Rantala, J. K. (2011). A cell spot microarray method for high-throughput biology: Dissertation. Turku: University of Turku.
Rantala, Juha K.. / A cell spot microarray method for high-throughput biology : Dissertation. Turku : University of Turku, 2011. 48 p.
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Rantala, JK 2011, 'A cell spot microarray method for high-throughput biology: Dissertation', Doctor Degree, University of Turku, Turku.

A cell spot microarray method for high-throughput biology : Dissertation. / Rantala, Juha K.

Turku : University of Turku, 2011. 48 p.

Research output: ThesisDissertationCollection of Articles

TY - THES

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AB - High-throughput screening of cellular effects of RNA interference (RNAi) libraries is now being increasingly applied to explore the role of genes in specific cell biological processes and disease states. However, the technology is still limited to specialty laboratories, due to the requirements for robotic infrastructure, access to expensive reagent libraries, expertise in high-throughput screening assay development, standardization, data analysis and applications. In the future, alternative screening platforms will be required to expand functional large-scale experiments to include more RNAi constructs, allow combinatorial loss-of-function analyses (e.g. genegene or gene-drug interaction), gain-of-function screens, multi-parametric phenotypic readouts or comparative analysis of many different cell types. Such comprehensive perturbation of gene networks in cells will require a major increase in the flexibility of the screening platforms, throughput and reduction of costs. As an alternative for the conventional multi-well based high-throughput screening -platforms, here the development of a novel cell spot microarray method for production of high density siRNA reverse transfection arrays is described. The cell spot microarray platform is distinguished from the majority of other transfection cell microarray techniques by the spatially confined array layout that allow highly parallel screening of large-scale RNAi reagent libraries with assays otherwise difficult or not applicable to high-throughput screening. This study depicts the development of the cell spot microarray method along with biological application examples of high-content immunofluorescence and phenotype based cancer cell biological analyses focusing on the regulation of prostate cancer cell growth, maintenance of genomic integrity in breast cancer cells, and functional analysis of integrin protein-protein interactions in situ.

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Rantala JK. A cell spot microarray method for high-throughput biology: Dissertation. Turku: University of Turku, 2011. 48 p.