title: Blocking Zebrafish MicroRNAs with Morpholinos.
abstract: Antisense morpholino oligonucleotides have been commonly used in zebrafish to inhibit mRNA function, either by inhibiting pre-mRNA splicing or by blocking translation initiation. Even with the advent of genome editing by CRISP/Cas9 technology, morpholinos provide a useful and rapid tool to knockdown gene expression. This is especially true when dealing with multiple alleles and large gene families where genetic redundancy can complicate knockout of all family members. miRNAs are small noncoding RNAs that are often encoded in gene families and can display extensive genetic redundancy. This redundancy, plus their small size which can limit targeting by CRISPR/Cas9, makes morpholino-based strategies particularly attractive for inhibition of miRNA function. We provide the rationale, background, and methods to inhibit miRNA function with antisense morpholinos during early development and in the adult retina in zebrafish.
authors.affiliation: Department of Biological Sciences, University of Southern Mississippi, Hattiesburg, MS, USA.
authors.forename: Alex Sutton
authors.initials: AS
authors.lastname: Flynt
authors.affiliation: Department of Biological Sciences, Vanderbilt University, 2325 Stevenson Center, Box 1820 Station B, Nashville, TN, 37235, USA.
authors.forename: Mahesh
authors.initials: M
authors.lastname: Rao
authors.affiliation: Department of Biological Sciences, Vanderbilt University, 2325 Stevenson Center, Box 1820 Station B, Nashville, TN, 37235, USA. James.G.Patton@Vanderbilt.edu.
authors.forename: James G
authors.initials: JG
authors.lastname: Patton
mesh_terms: D000818:Animals; D018274:Electroporation; D005786:Gene Expression Regulation; D020868:Gene Silencing; D018014:Gene Transfer Techniques; D035683:MicroRNAs / Q000235:genetics*; D008845:Microinjections; D060172:Morpholinos / Q000008:administration & dosage / Q000235:genetics*; D016376:Oligonucleotides, Antisense / Q000235:genetics*; D012160:Retina / Q000378:metabolism; D015027:Zebrafish / Q000235:genetics*
chemical_list: D035683:MicroRNAs; D060172:Morpholinos; D016376:Oligonucleotides, Antisense
keywords: Development; Embryo; Morpholino; Retina; Zebrafish; miRNA