Generation of Aβ-specific Tregs using CRISPR-Cas9

This experiment involved engineering regulatory T cells (Tregs) to express amyloid-beta (Aβ) specific T cell receptors (TCRAβ). The process used CRISPR-Cas9 technology to knock out the endogenous TCR in Tregs, followed by incorporation of a transgenic TCRAβ that was originally identified from Aβ-reactive effector T cell monoclones. The engineered TCRAβ-Tregs were validated for antigen specificity using MHC-Aβ-tetramer staining. This represents a novel approach to create disease-specific immunotherapy by conferring antigen specificity to naturally immunosuppressive Tregs, potentially allowing them to specifically target brain regions affected by Alzheimer's disease pathology while maintaining their regulatory functions.