Native homing endonucleases can target conserved genes in humans and in animal models.

Détails

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Etat: Public
Version: de l'auteur
ID Serval
serval:BIB_8BF5B419C126
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Native homing endonucleases can target conserved genes in humans and in animal models.
Périodique
Nucleic Acids Research
Auteur(s)
Barzel A., Privman E., Peeri M., Naor A., Shachar E., Burstein D., Lazary R., Gophna U., Pupko T., Kupiec M.
ISSN
1362-4962 (Electronic)
ISSN-L
0305-1048
Statut éditorial
Publié
Date de publication
2011
Peer-reviewed
Oui
Volume
39
Numéro
15
Pages
6646-6659
Langue
anglais
Résumé
In recent years, both homing endonucleases (HEases) and zinc-finger nucleases (ZFNs) have been engineered and selected for the targeting of desired human loci for gene therapy. However, enzyme engineering is lengthy and expensive and the off-target effect of the manufactured endonucleases is difficult to predict. Moreover, enzymes selected to cleave a human DNA locus may not cleave the homologous locus in the genome of animal models because of sequence divergence, thus hampering attempts to assess the in vivo efficacy and safety of any engineered enzyme prior to its application in human trials. Here, we show that naturally occurring HEases can be found, that cleave desirable human targets. Some of these enzymes are also shown to cleave the homologous sequence in the genome of animal models. In addition, the distribution of off-target effects may be more predictable for native HEases. Based on our experimental observations, we present the HomeBase algorithm, database and web server that allow a high-throughput computational search and assignment of HEases for the targeting of specific loci in the human and other genomes. We validate experimentally the predicted target specificity of candidate fungal, bacterial and archaeal HEases using cell free, yeast and archaeal assays.
Mots-clé
Algorithms, Animals, Archaea/enzymology, Bacteria/enzymology, Base Sequence, Conserved Sequence, Databases, Nucleic Acid, Endodeoxyribonucleases/genetics, Endodeoxyribonucleases/metabolism, Fungi/enzymology, Gene Targeting, Humans, Models, Animal, Mutation
Pubmed
Web of science
Open Access
Oui
Création de la notice
31/05/2011 9:02
Dernière modification de la notice
20/08/2019 14:50
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