Computational reprogramming of homing endonuclease specificity at multiple adjacent base pairs.

ABSTRACT: Site-specific homing endonucleases are capable of inducing gene conversion via homologous recombination. Reprogramming their cleavage specificities allows the targeting of specific biological sites for gene correction or conversion. We used computational protein design to alter the cleavage specificity of I-MsoI for three contiguous base pair substitutions, resulting in an endonuclease whose activity and specificity for its new site rival that of wild-type I-MsoI for the original site. Concerted design for all simultaneous substitutions was more successful than a modular approach against individual substitutions, highlighting the importance of context-dependent redesign and optimization of protein-DNA interactions. We then used computational design based on the crystal structure of the des

SUBMITTER: Ashworth J 

PROVIDER: S-EPMC2938204 | biostudies-literature | 2010 Sep

REPOSITORIES: biostudies-literature