Coordinated Edits: Simultaneously edit epistatically linked genes (e.g., opsin and ADCY8 for vision-cognition synergy, or angiopoietin with muscle-specific genes for circulatory-aerodynamic integration) to mimic rapid, coordinated evolution observed in Peregrine genomes.
Outcome Measures: Assess phenotypic outcomes in cell lines or embryonic models, focusing on functional proxies such as: (a) visual processing efficiency (e.g., simulated neural response to visual stimuli); (b) metabolic resilience (e.g., oxygen consumption rates under stress, mimicking stoop diving conditions); and (c) gene expression profiles to confirm epistatic or pleiotropic interactions. Expected results are that single-gene edits yield suboptimal or deleterious phenotypes, while coordinated edits enhance trait functionality, supporting the necessity of synchronized mutations.
Falsifiability Criteria
Null Hypothesis: Isolated mutations in genes like opsin or angiopoietin are sufficient to confer fitness advantages, consistent with gradual, partial evolution. If single-gene edits produce viable phenotypes (e.g., improved prey tracking or aerobic capacity without complementary changes), the rapid coordination hypothesis would be falsified.
Alternative Hypothesis: Coordinated mutations across epistatic and pleiotropic gene networks are required for functional adaptations, as isolated mutations result in non-viable intermediates (e.g., enhanced vision without cognitive processing fails to improve hunting success, currently 30--50% in Peregrines). If coordinated edits significantly outperform single-gene edits, this supports the rapid, synchronized evolution model driven by arms race pressures.
Validation Metrics: Use quantitative measures such as gene expression levels (via RNA-seq), protein interaction networks (via proteomics), and simulated fitness outcomes (e.g., cellular resilience to high-oxygen demands) to compare single vs. coordinated edits. Statistical tests (e.g., ANOVA, differential expression analysis) will assess significance of phenotypic differences.
Complementary In Silico Simulations
To complement CRISPR experiments, in silico simulations of gene networks will model epistatic and pleiotropic interactions using bioinformatics tools like STRING or Cytoscape, based on 2024--2025 genomic datasets. These simulations will: (a) predict fitness outcomes of single vs. coordinated mutations under arms race conditions (e.g., prey with zig-zag or flocking strategies); and (b) validate the rapid fixation of alleles in small populations, as observed in Peregrine subspecies post-bottlenecks (100,000--20,000 years ago). Simulations will incorporate ecological parameters, such as prey escape dynamics, to contextualize genetic findings.
These falsifiability tests provide a robust framework to evaluate the necessity of coordinated genetic changes in the Peregrine Falcon, ensuring that the proposed model of rapid, synchronized evolution is empirically testable and aligns with the species' ecological and genomic context.
IV. Results
A. Genomic Evidence: Rapid Selection in Migration/Adaptation Genes (ADCY8, BDNF); Low Diversity Enabling Quick Allele Fixation
