News

The RNAi Global Initiative convenes in Dublin: Focus on high-throughput and high-content methods for RNAi screens

Dublin, Ireland September 2008
Since its inception in 2005, the RNAi Global Initiative has met twice a year to review critical topics that have arisen during the course of their screening activities.  At the forefront of the seventh of these meetings held in Dublin (21 – 24 September 2008), discussions highlighted the integration of expanded and novel technologies into screening platforms, strategies and detection methods that yield high confidence candidate hits from distinct screens, and progress with developing experimental guidelines under MIARE, the minimum information guideline specific to RNAi experiments.  In addition, members reported on the outcomes of key collaborative activities facilitated by their participation in the RNAi Global Initiative. 

A question of particular interest to attendees in Dublin – many who are actively screening the whole human genome or focused collections (e.g., Kinome, druggable set, etc) – What are the best strategies to validate candidate gene targets derived from primary RNAi-based screens? The consensus was that while this bears further review, researchers have a variety of choices that include confirmation of phenotypes with independent reagents – including independent ON-TARGETplus™ SMARTpool® reagents enhanced for specificity, SMARTvector® shRNA lentiviral reagents for assessing long-term silencing, Accell™ siRNA reagents to investigate candidate targets in difficult to transfect but more relevant cell types, or non-RNAi reagents such as small molecule compounds.  In addition, they agreed that confirmation with alternative assays was important including rescreens in dominant-negative backgrounds or the application of high content approaches that permit multi-parametric analysis of distinct phenotypes (e.g., antibody-based (Cellomics HitKits™) or fluorescent-based (BioImage Redistribution® Assays).  The meeting closed with a well-received workshop dedicated to practical considerations for high content screening delivered by experts from the Thermo Fisher Scientific Cellomics products group. 

The RNAi Global Initiative - Work Group Activities
The RNAi Global Initiative represents a research alliance sponsored by Thermo Fisher Scientific that has a specific interest in sharing and promoting best practices and strategies for high throughput screening and high content analysis.  All members employ the siGENOME® whole-human genome of SMARTpool siRNA library (Thermo Scientific Dharmacon Products) which serves as one of their primary tools for pathway analysis or genome-wide silencing screens.  The outcomes of member screening efforts appear in key peer-reviewed publications.  (For a list of these peer-reviewed publications, please visit the RNAi Global website.)

The Initiative also supports focus group forums where members can address specific practical issues related to a particular area of discussion.  For example, the Informatics Work Group champions efforts towards establishing standardized reporting methods for RNAi-based screens using the Minimal Information About RNAi Experiments (MIARE) guidelines as the primary vehicle.  The broader scientific community is encouraged to take advantage of, and comment on the proposed standards, by visiting www.miare.org.  Further, the Informatics Work Group is collaborating with the National Center for Biotechnology Information (NCBI) to develop a means by which scientists can populate existing databases – Probe, Substance and BioAssay – with information about siRNAs.  The goal is to provide a public repository that includes annotation updates and links to researcher-generated data associated with specific silencing reagents (Probe and PubChem).   Yet another work group is focused on developing resource material for high content methods while a third work group actively promotes collaborations amongst members with related interests in infectious diseases, the products of which are expected to appear in up-coming publications.

The RNAi Global Initiative continues to serve as an example of a truly functional, global working group that actively fosters sharing best practices and collaborative activities.   For more information, please visit www.rnaiglobal.org.