SeQure DX™ Enabled Pre-Clinical Risk Assessment Of PCSK9 Guide RNAs

For the millions of people affected by ischemic heart disease, therapies that reduce cholesterol levels are critical. Gene editing offers a promising alternative to current treatments for high LDL cholesterol that require repeated dosing, as it has the potential for durable gene knockdown with a single delivery. In this case study, a series of guide RNAs (gRNAs) designed to reduce PCSK9 expression were analyzed to assess their on- and off-target editing risk.

Using a stepwise process, the gRNAs were screened to identify on-target variants and potential off-target editing sites. The top three performing gRNAs were then subjected to more detailed profiling, with the best candidate moving on to a final, high-sensitivity analysis. The results underscore the importance of considering genomic diversity in risk assessment, as most of the identified high-risk off-target sites would have been missed by screening the human reference genome alone.