CRISPR/Cas9 Disrupts T-Cell Receptor Expression
Gene editing nucleases can be used to disrupt normal T-cell receptor (TCR) expression, a desired effect when using T-cells as adoptive immunotherapy for cancer. This strategy targets the components of the TCR complex comprised of alpha and beta chains encoded by the TCRα (TRAC) or TCRb (TCRB) genes, respectively. Disrupting the gene sequence of either of these chains results in the ablation of the TCR complex and an inability of the cell to cause pathogenic graft versus host disease. A new method, utilizing the CRISPR/Cas9 platform, features the highest reported rates of TCR disruption by TRAC gene targeting.
Lower Risk of Graft Versus Host Disease
While adoptive immunotherapy shows promise in harnessing the anti-tumor effects of T-cells, the practice puts patients at risk for dangerous off-tumor anti-host (i.e., graft versus host disease) reactivity. In this new method, three classes of nuclease candidates that target the same region (exon 1) of the TRAC gene were assessed for TCRα gene disruption along with potential off-target cleavage. The CRISPR/Cas9 reagent showed the highest disruption efficiency with the lowest levels of toxicity and no off-target cleavage, making it the ideal candidate for disrupting TCR expression in a robust, scalable fashion with maximal safety.
BENEFITS AND FEATURES OF T-CELL RECEPTOR GENE EDITING AND LYMPHOCYTE ENGINEERING:
- Both megaTAL and CRISPR/Cas9 reagents are effective
- Effectively disrupts T-cell receptor expression
- The CRISPR/Cas9 reagent has low levels of toxicity and no off-target cleavage
- GMP compatible reagents and culture conditions represent a promising potential use as adoptive immunotherapy for cancer
Phase of Development In vitro