Massive sequencing projects expose the extent of natural, genetic diversity. Here, we describe a method with capacity to perform ancestor sequence reconstruction from data sets in excess of 10,000 sequences, poised to recover ancestral diversity, including the evolutionary events that determine present-time biological function and structure.
We introduce a novel strategy for suggesting “insertion-deletion variants” that are distinct from, but can be explored alongside, substitution variants for creating ancestral libraries. We demonstrate how insertions and deletions can be used as building blocks to form “hybrid ancestors”; based on this strategy, we synthesise ancestor variants, with varying enzymatic activities, for wide-ranging applications in the biotechnology sector.