Application Notes
Quantitative, single-cell analysis of siRNA treatment using smFISH
2025 년 05 월
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