AI Article Synopsis
- A new assay has been developed to measure mRNA knockdown caused by siRNA therapeutics across different animal species, offering significant time and cost savings in preclinical studies.
- The assay utilizes a sensitive stem-loop RT-qPCR technique that allows for quantification of specific mRNA without needing to purify RNA from tissue samples.
- This method could become a standard approach for assessing the effectiveness of siRNA treatments by providing a more straightforward and reliable means of measuring mRNA changes.
Article Abstract
Introduction: Various animal models are routinely used to evaluate the efficacy and toxicity of small interfering RNA (siRNA) therapeutics. Given that the most common measure of efficacy with siRNA therapeutics is mRNA knockdown, the development of a single assay for quantification of siRNA-mediated mRNA knockdown in multiple species would provide significant time and cost-savings during preclinical development.
Methods And Results: We have developed an assay targeting short consensus sequences of a particular mRNA in multiple species using the principles of a recently-reported stem-loop RT-qPCR method (Chen et al., 2005). The multi-species RT-qPCR assay is highly sensitive, reproducible, has a dynamic range of seven orders of magnitude, and it can be used to quantify a specific mRNA in crude tissue homogenates without the need for RNA purification. Compared to the limitations of conventional RT-qPCR assays, this assay provides a simple and robust tool for mRNA quantification to evaluate siRNA-mediated mRNA knockdown.
Discussion: This assay can potentially become a routine method for mRNA quantification to evaluate siRNA-mediated mRNA knockdown.
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| http://dx.doi.org/10.1016/j.vascn.2010.09.006 | DOI Listing |
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