ASO/siRNA Off-Target Safety Assessment

ASO/siRNA Off-Target Safety Assessment

Antisense oligonucleotides (ASO) and small interfering RNA (siRNA), as major categories of nucleic acid therapeutics, achieve gene silencing through sequence-specific binding to target mRNAs. However, unintended off-target effects may trigger immune activation, cytotoxicity, or non-target gene expression dysregulation, posing safety risks. GeneRulor provides comprehensive off-target detection and safety assessment services covering the full development lifecycle of nucleic acid drugs. At the target design stage, we apply bioinformatics algorithms for ASO/siRNA sequence design and optimization, predicting potential off-target genes by aligning against human transcriptome databases to select high-specificity candidate sequences. At the chemical synthesis stage, we offer ASO/siRNA synthesis services with multiple modification types (e.g., 2'-O-methyl, phosphorothioate, GalNAc conjugation), accompanied by in vitro screening and validation experiments to evaluate the effects of different sequences and modification strategies on target gene silencing efficiency and off-target activity. Computational off-target prediction tools combining seed-sequence matching, thermodynamic energy calculations, and conservation analysis systematically predict off-target risk genes. RNA-seq off-target detection, as the core technology, enables unbiased identification of unexpectedly downregulated or upregulated genes through whole-transcriptome sequencing comparison between treatment and control groups, revealing signal pathway perturbations and potential toxic responses caused by off-target effects. For key predicted off-target sites and differentially expressed genes, we provide RT-qPCR on-target/off-target validation services, ensuring data reliability through targeted quantitative confirmation of RNA-seq findings. The platform is applicable to ASO/siRNA products across multiple delivery strategies — liver-targeted, muscle-targeted, and CNS-targeted — supporting the off-target safety assessment data package required for IND submission and providing molecular mechanistic insights for dose optimization and toxicology studies during the clinical stage.