title: Targeting Functional Noncoding RNAs.
abstract: Noncoding RNAs have essential biochemical functions in different areas of cellular metabolism, including protein synthesis, RNA splicing, protein secretion, and DNA replication. We have successfully used Morpholino antisense oligonucleotides for the functional inactivation of small noncoding RNAs required for DNA replication (Y RNAs in vertebrates and stem-bulge RNAs in nematodes). Here we discuss specific issues of targeting functional noncoding RNAs for inactivation by Morpholino antisense oligonucleotides. We present protocols for the design, preparation, and efficacy controls of Morpholino antisense oligonucleotides, as well as brief descriptions for their delivery into vertebrate and nematode embryos.
authors.affiliation: Department of Zoology, University of Cambridge, Downing Street, Cambridge, CB2 3EJ, UK.|Department of Chemical and Systems Biology, Stanford University School of Medicine, Stanford, CA, 94305, USA.
authors.forename: Madzia P
authors.initials: MP
authors.lastname: Crossley
authors.affiliation: Department of Zoology, University of Cambridge, Downing Street, Cambridge, CB2 3EJ, UK. tk218@cam.ac.uk.
authors.forename: Torsten
authors.initials: T
authors.lastname: Krude
mesh_terms: D000818:Animals; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D018390:Gene Targeting; D018014:Gene Transfer Techniques; D060172:Morpholinos / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics*; D016376:Oligonucleotides, Antisense / Q000235:genetics; D010641:Phenotype; D022661:RNA, Untranslated / Q000235:genetics*; D013223:Statistics as Topic / Q000379:methods
chemical_list: D060172:Morpholinos; D016376:Oligonucleotides, Antisense; D022661:RNA, Untranslated
keywords: Caenorhabditis elegans; Danio rerio; Dominant-negative inhibition; Functional assay to test Morpholino efficacy and specificity; Functional inhibition; Morpholino delivery; Morpholino design; Noncoding RNA domains; Noncoding RNAs; Xenopus laevis