A novel framework for engineering protein loops exploring length and compositional variation.

ABSTRACT: Insertions and deletions (indels) are known to affect function, biophysical properties and substrate specificity of enzymes, and they play a central role in evolution. Despite such clear significance, this class of mutation remains an underexploited tool in protein engineering with few available platforms capable of systematically generating and analysing libraries of varying sequence composition and length. We present a novel DNA assembly platform (InDel assembly), based on cycles of endonuclease restriction digestion and ligation of standardised dsDNA building blocks, that can generate libraries exploring both composition and sequence length variation. In addition, we developed a framework to analyse the output of selection from InDel-generated libraries, combining next generation sequen

SUBMITTER: Tizei PAG 

PROVIDER: S-EPMC8080606 | biostudies-literature | 2021 Apr

REPOSITORIES: biostudies-literature