Engineering domain fusion chimeras from I-OnuI family LAGLIDADG homing endonucleases.

ABSTRACT: Although engineered LAGLIDADG homing endonucleases (LHEs) are finding increasing applications in biotechnology, their generation remains a challenging, industrial-scale process. As new single-chain LAGLIDADG nuclease scaffolds are identified, however, an alternative paradigm is emerging: identification of an LHE scaffold whose native cleavage site is a close match to a desired target sequence, followed by small-scale engineering to modestly refine recognition specificity. The application of this paradigm could be accelerated if methods were available for fusing N- and C-terminal domains from newly identified LHEs into chimeric enzymes with hybrid cleavage sites. Here we have analyzed the structural requirements for fusion of domains extracted from six single-chain I-OnuI family LHEs, spann

SUBMITTER: Baxter S 

PROVIDER: S-EPMC3439895 | biostudies-literature | 2012 Sep

REPOSITORIES: biostudies-literature