Molecular Recording Enables High-throughput Measurement of Biomolecular Activities
Methods of measuring the activities of biomolecules are fundamental to the study and engineering of biological systems. Fluorescence- or luminescence-based reporter assays are limited in their throughput, accuracy and dynamic range. Here we use a single long-read sequencing step to retrieve both sequence identity and biomolecular activity of each variant in a pooled library, by coupling the activity of interest to the targeted mutagenesis of a DNA sequence physically linked to the coding sequence of each variant. Our technique can potentially replace reporter-based flow cytometry activity assays and generate large datasets for mapping molecular fitness landscapes.