Fig. 2. Conception and validation of the TCR-CD3-MHC Class I immune synapse as a screening model for protein disruption.

a Diagram of the multimeric TCR-CD3 complex and MHC Class I immune synapse containing multiple spliced genes, based on the solved structures (PDB 6JXR⁴⁴, PDB 3T0E⁴⁵; PDB 10GA⁴⁷). b Diagram of the synthesis and localization of the TCR-CD3 complex and interaction with MHC Class I. All members of the CD3 complex are required before functional localization to the cell surface, where disruption of a single splice site within one gene member can prevent a surface expressed complex from forming. c Cas9 nuclease knockout of each individual member of TCR-CD3 complex validates the screening model. Two Cas9 nuclease sgRNAs were designed to exonic regions of each gene in the complex. All genes had at least one guide with ≥85% indel efficiency and loss in TCR-CD3 surface expression. Height of bars represents mean of N = 2 independent donors. Source data are available in the Source Data file.