Deliver overexpression/knockdown vectors via viral packaging, or directly knock out the target gene via gene editing, to establish a cell/animal model with altered expression.
Observe behavioral-level changes at the cellular level after the expression shift, via proliferation, apoptosis, migration, and invasion assays.
Validate mRNA-level changes with qPCR and protein-level changes with Western Blot, confirming the causal relationship between the phenotype and the target gene at the molecular level.
Combine the phenotypic and molecular-level evidence to establish whether Gene X promotes or inhibits the biological process, and the underlying causal relationship.
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