The most informative design is a layered perturbation experiment in human neurons exposed to proteasome or ER stress: paired nascent-RNA profiling, ChIP/CUT&RUN for candidate factors, and CRISPR perturbations of ATF4, DDIT3, HSF1, and REST. The goal is to distinguish direct promoter/enhancer occupancy from indirect network-level repression.
ATF4/DDIT3 ranked highest because they are central stress integrators and offer a plausible route to a reversible tau-lowering response. HSF1 remains compelling because it links proteostasis rescue to transcriptional reprioritization, but a targeted repressor role is not yet proven. REST should be treated as a stage-specific branch that could matter in chronic or severe stress states rather than as the default first mechanism.