GROQ-seq Technical Status Report January 2025

This technical report outlines the first-year progress of the GROQ-seq initiative—a high-throughput platform for quantitative protein function assays based on growth-coupled genetic circuits in Escherichia coli (E. coli). Developed collaboratively by The Align Foundation and NIST, GROQ-seq enables sequence-to-function mapping for engineered proteins at scale, supporting synthetic biology and machine learning efforts in protein design.

Key innovations include:

• Replacement of TetA with dihydrofolate reductase (DHFR)-based circuits, achieving >100× dynamic range
• Modular plasmid design using Golden Gate assembly
• Integration of inducible promoters and standardized Teknova media to improve reproducibility across biofoundry-style workflows
• Onboarding and validation of seven function-specific genetic assays (e.g., for transcription factors, proteases, tRNA synthetases)

These standardized, growth-based assays support large-scale generation of genotype-phenotype datasets, enabling predictive models of protein function. This work advances industrial molecular engineering of proteins, synthetic biology platforms, and collaborative data-sharing infrastructure.

10.6084/m9.figshare.28963640