Crystallographic analyses illustrate significant plasticity and efficient recoding of meganuclease target specificity.

ABSTRACT: The retargeting of protein-DNA specificity, outside of extremely modular DNA binding proteins such as TAL effectors, has generally proved to be quite challenging. Here, we describe structural analyses of five different extensively retargeted variants of a single homing endonuclease, that have been shown to function efficiently in ex vivo and in vivo applications. The redesigned proteins harbor mutations at up to 53 residues (18%) of their amino acid sequence, primarily distributed across the DNA binding surface, making them among the most significantly reengineered ligand-binding proteins to date. Specificity is derived from the combined contributions of DNA-contacting residues and of neighboring residues that influence local structural organization. Changes in specificity are facilitated

SUBMITTER: Werther R 

PROVIDER: S-EPMC5737575 | biostudies-literature | 2017 Aug

REPOSITORIES: biostudies-literature