Insect-based circadian and sleep screening platform for botanical bioactives

Overview

This platform provides an integrated, non-animal in vivo screening service for evaluating botanical extracts and dietary bioactives targeting sleep and circadian health. Using synchronized mosquito cohorts, the system delivers dual-endpoint readouts: high-resolution molecular clock profiling through timeless qPCR dynamics and RNA-seq, alongside automated video-tracked behavioral sleep measurements including latency, total duration, and consolidation.

The whole-organism approach captures integrated transcriptomic responses across digestive, metabolic, and neural pathways in a single assay. This offers a scalable, cost-effective screening solution that bridges the translational gap between simplified cell-based assays and costly human clinical trials, enabling faster and more informed decisions on bioactive ingredient development.

Technical specifications

Core methodology:

• Synchronized cohorts of the strictly nocturnal mosquito Anopheles dirus maintained under controlled light-dark cycles (LD 12:12), with 10 females per 6-oz cup
• Oral administration of test ingredients through a 10% sucrose liquid matrix, enabling homogeneous dosing of complex botanical extracts without altering food texture or palatability
• Molecular clock profiling via timeless qPCR dynamics across multiple circadian timepoints (ZT21, ZT00, ZT03, ZT09, ZT12, ZT15)
• Whole-organism RNA-seq for transcriptomic pathway mapping across digestive, metabolic, and neural systems using GO/KEGG analysis
• Automated IR video tracking for behavioral sleep quantification, defined as continuous immobility of 5 minutes or longer

Key advantages:

• Nocturnal vector biology directly aligns with nighttime sleep physiology, providing high sensitivity to sleep-promoting compounds
• Simultaneous molecular and behavioral endpoints in a single experimental system
• Significantly lower cost than organoid-based approaches while delivering whole-organism integration that cell lines cannot provide

Technology readiness level

The platform is currently at TRL 3, with validated proof-of-concept data demonstrating high-sensitivity 24-hour timeless clock gene expression dynamics in synchronized cohorts. A phased validation roadmap is in place: Phase 1 focuses on infrastructure setup and calibration using melatonin and GABA reference controls; Phase 2 conducts tier 1 bioactive screening across concentrations to identify lead candidates; Phase 3 applies tier 2 mechanistic transcriptomics to top hits, delivering executive screening reports to support sponsor product development decisions.