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Genome-Wide RNAi Global Initiative

High-throughput RNAi loss-of-function screening in mammalian cells has become a valuable method to identify and describe genetic relationships in both basic biology and disease mechanisms.

See: Technology RNAi primer

Mission

The RNAi Global Initiative is an alliance of leading international research centers under the 09:02:36 AM Wed, Sep 08 2010auspices of Dharmacon, Inc. Its goal is to combine the powerful technology of genome-wide RNAi screening with international scientific data exchange in order to accelerate and facilitate basic biological and medical discovery.

Challenges

  • Unambiguous interpretation of results
  • Reproduction of results
  • Sharing of data
  • Comparison of data

Approach

Building on established standards such as MIAME (Minimum Information About a Microarray Experiment), the RNAi Global Initiative has contributed work towards a community-wide effort known as the Minimum Information About an RNAi Experiment (MIARE). MIARE was initiated to develop universal reporting standards and guidelines that would facilitate the exchange, analysis and annotation of data derived from RNAi-based screens.

To facilitate the development of the MIARE guidelines, ten member laboratories of RNAi Global conducted a multi-center siRNA benchmark screening experiment targeting 859 genes (779 genes of the phylogenetically related protein kinase family and 80 cell cycle genes) using two common assays: CellTiter-Blue® (Promega) and Apo-ONE® Homogeneous Caspase-3/7 (Promega).

Study design

Design of benchmarking study

Reporting Guidelines

MIARE is intended to be modular so that it may be coupled or readily integrated with related standards such as MIACA (Minimum Information About Cellular Assay). Below are MIARE sections that need to be described while reporting an experiment. Detailed specifications for each section can be found at http://www.miare.org.

  • Experiment description (goal, publication, contacts, etc.)
  • Experimental design (replicates, assays, etc.)
  • Sample description (bio-source description, treatments, etc.)
  • Assay plate description
  • Silencing RNA reagent (siRNAs, mixture components)
  • Delivery (transfection reagent, protocol, etc.)
  • Assay plate (conditions within the plate - time points, medium, etc.)
  • Post treatment
  • Assay (description, reagents, protocol, instruments, etc.)
  • Data analysis (software, normalization, scoring, etc.)
  • Data (quantitative and qualitative data, raw and processed data)

Resources

References

siRNA siRNA pool
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Last updated: 2014-09-24T14:40:50-04:00