An In Vivo Selection System for Homing Endonuclease Activity

Gruen, MathiasChang, KathySerbanescu, IrinaLiu, David2009-05-202002Gruen, Mathias, Kathy Chang, Irina Serbanescu, and David R. Liu. 2002. An In Vivo Selection System for Homing Endonuclease Activity. Nucleic Acids Research 30, no. 7: e29.0305-1048http://nrs.harvard.edu/urn-3:HUL.InstRepos:2961939Homing endonucleases are enzymes that catalyze the highly sequence-specific cleavage of DNA. We have developed an in vivo selection in Escherichia coli that links cell survival with homing endonuclease-mediated DNA cleavage activity and sequence specificity. Using this selection, wild-type and mutant variants of three homing endonucleases were characterized without requiring protein purification and in vitro analysis. This selection system may facilitate the study of sequence-specific DNA cleaving enzymes, and selections based on this work may enable the evolution of homing endonucleases with novel activities or specificities.en-USAn In Vivo Selection System for Homing Endonuclease Activity10.1093/nar/30.7.e29