Mining endonuclease cleavage determinants in genomic sequence data.

J Biol Chem

Department of Biochemistry, Howard Hughes Medical Institute, University of Washington, Seattle, Washington 98195, USA.

Published: September 2011

Homing endonucleases have great potential as tools for targeted gene therapy and gene correction, but identifying variants of these enzymes capable of cleaving specific DNA targets of interest is necessary before the widespread use of such technologies is possible. We identified homologues of the LAGLIDADG homing endonuclease I-AniI and their putative target insertion sites by BLAST searches followed by examination of the sequences of the flanking genomic regions. Amino acid substitutions in these homologues that were located close to the target site DNA, and thus potentially conferring differences in target specificity, were grafted onto the I-AniI scaffold. Many of these grafts exhibited novel and unexpected specificities. These findings show that the information present in genomic data can be exploited for endonuclease specificity redesign.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3173205PMC
http://dx.doi.org/10.1074/jbc.M111.259572DOI Listing

Publication Analysis

Top Keywords

mining endonuclease
4
endonuclease cleavage
4
cleavage determinants
4
determinants genomic
4
genomic sequence
4
sequence data
4
data homing
4
homing endonucleases
4
endonucleases great
4
great potential
4

Similar Publications

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!