Fig. 1
From: Amyloid-β specific regulatory T cells attenuate Alzheimer’s disease pathobiology in APP/PS1 mice
Generation of TCRAβ-Tregs. A. Flow cytometry gating confirming the CRISPR-Cas9-mediated knockout of endogenous T cell receptors (TCRs) to generate TCR−−-Tregs. Nucleotide sequences represent the guide RNAs targeting the alpha and beta regions of the TCR. B. Plasmid design for TCRAβ electroporation. C. Representative gating strategy confirming the TCR expression by TCR-Tregs electroporated with 0.5 μL of TCRAβ plasmid (plasmid conc. = 1.5 μg/μL). D. Time course of TCRAβ expression on engineered TCRAβ-Tregs post electroporation with TCRAβ plasmid. Phenotype characterization of engineered TCRAβ-Tregs shown in Supplementary Fig. 3. E. MHCII-IA.b-KLVFFAEDVGSNKGA (Aβ T cell epitope) tetramer binding confirming the Aβ reactivity of engineered TCRAβ-Tregs incubated with Aβ-tetramer (blue) compared to TCRAβ-Tregs incubated with control-tetramer (orange) or polyclonal Tregs incubated with Aβ-Tetramer (red)