T-cell genomic DNA at engineered CRISPR loci

T-cell genomic DNA at engineered CRISPR loci refers to specific sequences within the T-cell genome targeted for modification using CRISPR-Cas technology to enhance therapeutic potential. This target is central to the production of next-generation adoptive cell therapies, such as CAR-T cells, where specific loci like the T-cell receptor alpha constant (TRAC) are edited to prevent graft-versus-host disease in allogeneic settings (Eyquem et al., 2017, Nature). By utilizing guide RNAs to direct Cas nucleases, precise double-strand breaks are introduced, allowing for the knockout of inhibitory receptors like PD-1 or the site-specific insertion of therapeutic transgenes (Stadtmauer et al., 2020, Science). Unlike traditional viral transduction, targeting specific genomic loci allows for more uniform transgene expression and avoids the risks associated with semi-random integration (Roth et al., 2018, Nature). However, the use of genomic DNA as a target requires careful evaluation of off-target editing and potential chromosomal translocations that could lead to oncogenic transformation (Kosicki et al., 2018, Nature Biotechnology). Clinical applications currently focus on oncology and infectious diseases, where precise genome editing can significantly improve the persistence and potency of engineered T-cells (June et al., 2018, Science). The interaction between the CRISPR-Cas complex and the DNA target is governed by the protospacer adjacent motif (PAM) and sequence complementarity of the guide RNA (Jinek et al., 2012, Science).

Other names

T-cell genomeCRISPR-modified T-cell DNAEngineered T-cell genetic lociTRAC locus (engineered)PDCD1 locus (engineered)

Targeted genomic disruption via Cas-mediated double-strand breaks followed by non-homologous end joining (NHEJ) or homology-directed repair (HDR).