title: Targeting human microRNA genes using engineered Tal-effector nucleases (TALENs).
abstract: MicroRNAs (miRNAs) have quickly emerged as important regulators of mammalian physiology owing to their precise control over the expression of critical protein coding genes. Despite significant progress in our understanding of how miRNAs function in mice, there remains a fundamental need to be able to target and edit miRNA genes in the human genome. Here, we report a novel approach to disrupting human miRNA genes ex vivo using engineered TAL-effector (TALE) proteins to function as nucleases (TALENs) that specifically target and disrupt human miRNA genes. We demonstrate that functional TALEN pairs can be designed to enable disruption of miRNA seed regions, or removal of entire hairpin sequences, and use this approach to successfully target several physiologically relevant human miRNAs including miR-155*, miR-155, miR-146a and miR-125b. This technology will allow for a substantially improved capacity to study the regulation and function of miRNAs in human cells, and could be developed into a strategic means by which miRNAs can be targeted therapeutically during human disease.
authors.affiliation: Department of Pathology, University of Utah, Salt Lake City, Utah, United States of America.
authors.forename: Ruozhen
authors.initials: R
authors.lastname: Hu
authors.forename: Jared
authors.initials: J
authors.lastname: Wallace
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Dahlem
authors.forename: David Jonah
authors.initials: DJ
authors.lastname: Grunwald
authors.forename: Ryan M
authors.initials: RM
authors.lastname: O'Connell
mesh_terms: D000483:Alleles; D000818:Animals; D001483:Base Sequence; D004720:Endonucleases / Q000378:metabolism*; D005818:Genetic Engineering; D056426:Genetic Loci / Q000235:genetics; D015894:Genome, Human / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D035683:MicroRNAs / Q000235:genetics*; D008969:Molecular Sequence Data; D059645:Mutation Rate; D009690:Nucleic Acid Conformation; D017384:Sequence Deletion / Q000235:genetics; D014162:Transfection
chemical_list: D035683:MicroRNAs; D004720:Endonucleases