title: An integrated miRNA functional screening and target validation method for organ morphogenesis.
abstract: The relative ease of identifying microRNAs and their increasing recognition as important regulators of organogenesis motivate the development of methods to efficiently assess microRNA function during organ morphogenesis. In this context, embryonic organ explants provide a reliable and reproducible system that recapitulates some of the important early morphogenetic processes during organ development. Here we present a method to target microRNA function in explanted mouse embryonic organs. Our method combines the use of peptide-based nanoparticles to transfect specific microRNA inhibitors or activators into embryonic organ explants, with a microRNA pulldown assay that allows direct identification of microRNA targets. This method provides effective assessment of microRNA function during organ morphogenesis, allows prioritization of multiple microRNAs in parallel for subsequent genetic approaches, and can be applied to a variety of embryonic organs.
authors.affiliation: Division of Genetics, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Ivan T
authors.initials: IT
authors.lastname: Rebustini
authors.affiliation: Division of Genetics, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Maryann
authors.initials: M
authors.lastname: Vlahos
authors.affiliation: Department of Molecular and Cell Biology, Boston University Medical Center, Boston, MA 02118, USA.
authors.forename: Trevor
authors.initials: T
authors.lastname: Packer
authors.affiliation: Department of Molecular and Cell Biology, Boston University Medical Center, Boston, MA 02118, USA.
authors.forename: Maria A
authors.initials: MA
authors.lastname: Kukuruzinska
authors.affiliation: Division of Genetics, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Richard L
authors.initials: RL
authors.lastname: Maas
mesh_terms: D000818:Animals; D005260:Female; D015870:Gene Expression; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D053263:Gene Regulatory Networks; D051379:Mice; D035683:MicroRNAs / Q000502:physiology*; D009928:Organ Specificity; D038081:Organogenesis; D034622:RNA Interference
chemical_list: D035683:MicroRNAs