Characterization of homing endonuclease binding and cleavage specificities using yeast surface display SELEX (YSD-SELEX).

ABSTRACT: LAGLIDADG homing endonucleases (LHEs) are a class of rare-cleaving nucleases that possess several unique attributes for genome engineering applications. An important approach for advancing LHE technology is the generation of a library of design ‘starting points’ through the discovery and characterization of natural LHEs with diverse specificities. However, while identification of natural LHE proteins by sequence homology from genomic and metagenomic sequence databases is straightforward, prediction of corresponding target sequences from genomic data remains challenging. Here, we describe a general approach that we developed to circumvent this issue that combines two technologies: yeast surface display (YSD) of LHEs and systematic evolution of ligands via exponential enrichment (SELEX). Usi

SUBMITTER: Jacoby K 

PROVIDER: S-EPMC5388424 | biostudies-literature | 2017 Feb

REPOSITORIES: biostudies-literature