Quantification of On/Off-Target Gene Editing

Quantitative analysis of gene editing is now an essential step in getting regulatory approval of gene editing-based therapies. But designing and validating assays for on/off-target editing analysis and building an internal team to perform them can be costly and inefficient.

Sensitive and high-throughput amplicon sequencing can accurately quantify on-target editing efficiency and off-target editing events at predicted loci, including chromosome translocation events. These analyses require an experienced team that operates in compliance with both GLP and GMP guidelines, to produce data that holds up against regulatory scrutiny.

To meet evolving regulatory requirements, standards and to mitigate risk, researchers utilizing tools such as CRISPR and other gene editing platforms must proactively identify and address off target effects, as well as any unintended modifications at a non-target site such as translocation possibilities.

Fortunately, modern genomic assay platforms — including NGS and ddPCR— provide effective, reliable means to identify off target effects and events, including point mutations, deletions, insertions, inversions, and unexpected translocations.

On-Target Gene Editing Quantification

IDT’s high-specificity rhAmpSeq™ amplicon sequencing system coupled with the Illumina next-gen sequencing platform produces deep insights into your on-target editing efficiency. The sensitivity of these assays enables the identification and characterization of extremely rare variants.

Off-Target Gene Editing Analysis

Using bioinformatics, one can predict the number of potential off-target sites and design custom assays that can analyze hundreds of loci using a single rhAmpSeq™ panel. This allows you to quickly and accurately sequence multiple confirmed or putative edit sites. One can also take a genome-wide, unbiased approach with sophisticated sequencing techniques.

On-Target Gene Editing Analysis with ddPCR

For additional editing quantification at specific target sequences, ddPCR can provide ultra-sensitive detection of translocation events. Bio-Rad’s Q200 ddPCR platform provides a cost-effective and accurate assessment of translocation events that may occur during gene editing.

Improve Off Target Analysis with Avance as Your Partner

As a true collaborative partner, Avance Biosciences™ helps you create a scientific and operational roadmap that helps you meet research, clinical, and commercial objectives. We’re ready and able to help you characterize your product’s off target site impact in detail, as well as navigate the evolving CRISPR gene editing standards laid out by the FDA and other regulatory bodies worldwide.