Biological part characterization is the core requirement for engineering complex, yet predictable biosystems. The immense complexity of nature makes this a challenging task. Currently, there is a considerable lack of well-defined, standardized parts and an insufficient grasp of their sequence-function relationship. Notably, state of the art screening methods have insufficient throughput to effectively navigate the extensive biomolecule sequence space. To address this issue we have developed a novel approach to part characterization based on microfluidics and modified nucleotides: Catalytic Activity Sequencing (CAT-Seq). CAT-Seq enables the simultaneous activity measurements of billions of biomolecule variants in parallel. Unique biomolecules are each synthesized in separate water droplets and their activity is recorded and stored into their individual DNA sequences. This information can then be readily retrieved by next-generation sequencing. CAT-Seq can rapidly assess sequence-function relationships, characterize regulatory parts, their interactions, and provide much-needed data for predictively designing novel biological systems.

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