UMI-linked consensus sequencing enables phylogenetic analysis of directed evolution

Florian Hollfelder, Paul Zurek, Ahir Pushpanath, Katharina Neufeld & Philip Knyphausen
The success of protein evolution campaigns is strongly dependent on the sequence context in which mutations are introduced, stemming from pervasive non-additive interactions between a protein’s amino acids (‘intra-gene epistasis’). Our limited understanding of such epistasis hinders the correct prediction of the functional contributions and adaptive potential of mutations. Here we present a straightforward unique molecular identifier (UMI)-linked consensus sequencing workflow (UMIC-seq) that simplifies mapping of evolutionary trajectories from full-length sequences. Attaching UMIs to gene...
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