title: MicroRNA in situ hybridization on whole-mount preimplantation embryos.
abstract: Whole-mount in situ hybridization (WISH) using antisense probes is widely used to visualize RNA sequences in embryos and to determine the precise site of expression in the different cells or tissues. The target sequence is hybridized with an antisense RNA probe, followed by visual or fluorescence detection to measure the site and level of expression. However, the detection of short RNA molecules is hampered by the reduced stringency of the probes for short transcripts. Here, we describe a procedure for WISH detection of short RNA molecules, like miRNAs, in mammalian preimplantation embryos using LNA-modified probes with high sensitivity and specificity.
authors.affiliation: Department of Nutrition, Genetics and Ethologyy - Faculty of Veterinary Medicine, Ghent University, Heidestraat 19, 9820, Merelbeke, Belgium, Karen.Goossens@UGent.be.
authors.forename: Karen
authors.initials: K
authors.lastname: Goossens
authors.forename: Luc
authors.initials: L
authors.lastname: Peelman
authors.forename: Ann
authors.initials: A
authors.lastname: Van Soom
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000378:metabolism* / Q000648:ultrastructure; D002417:Cattle; D018507:Gene Expression Regulation, Developmental; D017403:In Situ Hybridization / Q000379:methods*; D035683:MicroRNAs / Q000032:analysis* / Q000235:genetics; D009841:Oligonucleotides / Q000032:analysis / Q000235:genetics; D015347:RNA Probes / Q000032:analysis / Q000235:genetics; D016707:Tissue Fixation / Q000379:methods
chemical_list: D035683:MicroRNAs; D009841:Oligonucleotides; D015347:RNA Probes; C477371:locked nucleic acid