On-/Off-Target Analysis

Genome editing using designer nucleases such as CRISPR/Cas9, ZFNs and TALENs allow the targeted introduction of genetic material into a specific locus of the mammalian genome. However, undesired on- and off-target genome modifications may arise which could cause genomic instability and disrupt the functionality of otherwise normal genes potentially causing safety concerns in pre-clinical and clinical studies

Genome editing

Current off-target analysis techniques rely mainly on bioinformatics predictions and genome-wide off-target detection techniques. Such predictions lack reliability as they cover only a small portion of potential genomic alterations. And off-target mutations with a frequency below 0.5% remain mostly undetected by current genome-wide off-target detection techniques. We are the pioneer to develop unbiased assays for genome-wide on-/off-target analysis.

Targeted genome editing – delineating desired from undesired outcomes 

At GeneWerk, we offer comprehensive PCR and NGS-based assays for gene editing safety assessment. Our versatile and cost-effective assays interrogate both on- and off-target genomic alterations.

Our state-of-the-art assays in combination with powerful in-house bioinformatics pipelines reliably quantify the ratios between the desired targeted integration and non-desired outcomes such as translocations and indels.

PCR-Based

At GeneWerk, we adapted the Shearing Extension Primer Tag Selection Ligation-Mediated PCR (S-EPTS/LM PCR) method to analyze on-target and off-target integrations arising from genome editing.

  • Convenient detection of on-/off-target alterations and random integrations
  • Adaptable to address specific questions raised by regulatory authorities
  • Customized bioinformatics analysis
Read more about PCR

NGS-Based

At GeneWerk, we use target enrichment sequencing (TES) for genome-wide detection and quantification of on- and off-target alterations. Our time and cost-effective approach captures all possible on-/off-target events in a single reaction.

 

  • NGS-based approach – avoid PCR artifacts
  • Captures predicted as well as non-predicted on-/off-target events
  • Customized bioinformatics analysis
Read more about NGS

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