Notas de aplicação
Quantitative, single-cell analysis of siRNA treatment using smFISH
maio 2025
Nucleic acid therapeutics are innovative drugs composed of DNA or RNA approximately 18–30 nucleotides in length. They function by suppressing, modifying, or complementing the expression of disease-associated genes. In particular, siRNA therapeutics function by suppressing gene expression through sequence-specific degradation of target mRNA. Improving their pharmacokinetics and pharmacodynamics remains a critical design consideration. During discovery and screening stages, both efficacy and delivery efficiency at the cellular level are often used as key indicators and are commonly assessed using real-time PCR. However, real-time PCR encounters technical limitations in distinguishing sequence-dependent gene silencing efficacy from cellular delivery efficiency, which hinders accurate performance evaluation. This application note introduces a novel method utilizing smFISH (single-molecule fluorescence in situ hybridization), which enables RNA visualization at the single-molecule level, to distinctly report the knockdown efficacy and delivery efficiency of an siRNA treatment.
Keywords: smFISH, RNA imaging, General Analysis 3, siRNA therapeutics