Methods combine Agilent's bioanalyser with Qiagen's sample preparation and RNAi products to streamline experiments for drug discovery
Agilent Technologies has announced that it has introduced new methods for optimising RNAi experiments in gene silencing, combining the proprietary siRNA technology of Qiagen with Agilent's 2100 bioanalyser.
RNA interference (RNAi) is a research technique used in mammalian cell cultures to reduce the expression of selected genes in order to understand the role of their corresponding proteins.
RNAi technology is expected to have a significant impact on target validation and basic disease research, thereby helping to speed drug discovery.
In RNAi applications, small interfering RNAs (siRNAs) can be introduced (transfected) into cells in order to block or 'silence' the function of selected genes.
To be successful, RNAi scientists need an easy and effective way to optimise conditions for transfection, verify the quality of their siRNA, and measure the success of transfection.
Optimal transfection conditions vary significantly between cell lines.
Agilent and Qiagen have combined their technologies to create a fast, new solution for this application, using the Agilent 2100 bioanalyser with cell fluorescence LabChip kit, DNA 1000 LabChip kit, and the Qiagen TransMessenger and RNAiFect transfection reagents and fluorescently tagged siRNAs, which are based on Qiagen's patented Tom-Amidites synthesis chemistry.
Agilent's application note (publication 5988-9872EN), which describes this method, is available online from Agilent's Web site.
The Agilent 2100 bioanalyser combines electrophoretic analysis of nucleic acids with dual-colour flow cytometry capabilities. Compared to fluorescence microscopy, the Agilent 2100 bioanalyser automates measurements and provides quantitative information on the transfection degree and viability of each cell.
Compared to conventional flow cytometer instruments, this system provides substantial cost savings and is far easier to use.
Agilent says the 2100 bioanalyser is already established as the industry standard for RNA QC in gene expression studies, and this new application makes it a valuable analysis tool for gene silencing researchers.
Agilent Technologies and Qiagen are working together in additional areas using microfluidics to provide better solutions for gene silencing experiments.
Further applications using the Agilent 2100 bioanalyser and Qiagen reagents to monitor the silencing effect of RNAi by antibody staining of cells or analysis of endpoint RT-PCR results are expected to be available shortly.
The role of RNAi in drug target validation.
Most drugs target a specific protein, eliminating or changing its activity in order to improve the health of a patient.
In identifying proteins that can serve as drug targets, pharmaceutical and biotech companies go through an extensive process called target validation to verify that they understand the protein's function and its role in related pathways.
Frequently, this involves the breeding of 'knockouts', usually a strain of mouse in which the protein has been eliminated.
This process can take a year or more.
The introduction of RNAi into a cell, tissue culture or animal can potentially provide the same information as a knockout experiment at a lower cost and in less time.
