Above Fig.: Custom dGH in-Site™ probe set targeting TRAC (red), B2M (yellow) and CIITA (green) loci in a GM cell line.

The TRAC, CIITA and B2M genes are high-value editing targets in CAR-T cell engineering. Disabling them goes far towards mitigating the risk of graft versus host disease (GvHD) and towards ensuring modified T cells survive in a patient’s body long enough to reduce their tumor burden. However, editing methods are imperfect and can trigger unanticipated structural rearrangements at the edit sites. Some of these changes can be challenging to detect, especially when they result in no gain or loss of chromosomal material, or when they involve repetitive DNA.

Above Fig.: dGH in-Site™ probes for TRAC (orange), B2M (green) and CIITA (red) on GM14907 control lymphoblastoid cell line.

High-Value Editing Targets in CAR-T Cell Engineering.

The TRAC, CIITA and B2M genes are high-value editing targets in CAR-T cell engineering. Disabling them goes far towards mitigating the risk of graft versus host disease (GvHD) and towards ensuring modified T cells survive in a patient’s body long enough to reduce their tumor burden. However, editing methods are imperfect and can trigger unanticipated structural rearrangements at the edit sites. Some of these changes can be challenging to detect, especially when they result in no gain or loss of chromosomal material, or when they involve repetitive DNA.

dGH in-Site technology provides high-resolution data on the outcome of editing experiments with tailor-made TRAC, CIITA and B2M probes. These assay components can reveal rearrangements ranging from single-digit kilobase inversions, to translocations and deletions. Data supporting the safety profile of a therapy product needs to stand up to high regulatory standards, and dGH technology can provide a comprehensive picture of editing outcomes which other assays simply cannot.