Gene-Editing Off-Target Detection

Gene-Editing Off-Target Detection

As a cornerstone tool of precision medicine, the safety of gene-editing technologies hinges on the accurate identification and evaluation of off-target effects. GeneRulor has built a comprehensive off-target detection platform covering multiple editing systems — including CRISPR, base editors (ABE/CBE), and prime editors (PE) — spanning computational prediction, in vitro validation, and in vivo monitoring to form a complete risk assessment pipeline. We utilize professional tools such as CRISPRme for sgRNA design optimization and off-target site prediction, and employ cutting-edge unbiased whole-genome off-target capture technologies including GUIDE-seq, AID-seq, Detect-seq, inoSeq, and PE-tag. Results are cross-validated through WGS, whole-exome sequencing, and transcriptome sequencing. Additionally, we have developed liquid-phase hybridization capture technology for targeted deep sequencing verification of off-target sites, complemented by Sanger sequencing and amplicon sequencing. For iPSC and other stem cell products, we provide exome- and genome-level editing safety assessments to ensure genetic stability. The platform also offers process residual testing — including ELISA for Cas9 protein residuals and RT-qPCR for sgRNA residuals — as well as HLA typing (Sanger and third-generation sequencing), fully meeting regulatory requirements for allogeneic cell therapy products. From early-stage sgRNA screening to comprehensive off-target assessment for IND submission, GeneRulor delivers end-to-end solutions for the safety of gene-editing products.