title: Knockdown of αIIb by RNA degradation by delivering deoxyoligonucleotides piggybacked with control vivo-morpholinos into zebrafish thrombocytes.
abstract: Morpholino and vivo-morpholino gene knockdown methods have been used to study thrombocyte function in zebrafish. However, a large-scale knockdown of the entire zebrafish genome using these technologies to study thrombocyte function is prohibitively expensive. We have developed an inexpensive gene knockdown method, which uses a hybrid of a control vivo-morpholino and a standard antisense oligonucleotide specific for a gene. This hybrid molecule is able to deliver antisense deoxyoligonucleotides into zebrafish thrombocytes because it piggybacks on a control vivo-morpholino. To validate use of this hybrid molecule in gene knockdowns, we targeted the thrombocyte specific αIIb gene with a hybrid of a control vivo-morpholino and an oligonucleotide antisense to αIIb mRNA. The use of this piggyback technology resulted in degradation of αIIb mRNA and led to thrombocyte functional defect. This piggyback method to knockdown genes is inexpensive since one control vivo-morpholino can be used to target many different genes by making many independent gene-specific oligonucleotide hybrids. Thus, this novel piggyback technology can be utilized for cost-effective large-scale knockdowns of genes to study thrombocyte function in zebrafish.
authors.affiliation: Department of Biological Sciences, University of North Texas, Denton, TX 76203, USA.
authors.forename: Hemalatha
authors.initials: H
authors.lastname: Sundaramoorthi
authors.affiliation: Department of Biological Sciences, University of North Texas, Denton, TX 76203, USA.
authors.forename: Gauri
authors.initials: G
authors.lastname: Khandekar
authors.affiliation: Department of Biological Sciences, University of North Texas, Denton, TX 76203, USA.
authors.forename: Seongcheol
authors.initials: S
authors.lastname: Kim
authors.affiliation: Department of Biological Sciences, University of North Texas, Denton, TX 76203, USA. Electronic address: jag@UNT.edu.
authors.forename: Pudur
authors.initials: P
authors.lastname: Jagadeeswaran
mesh_terms: D000818:Animals; D001792:Blood Platelets / Q000378:metabolism*; D055785:Gene Knockdown Techniques / Q000379:methods*; D060172:Morpholinos / Q000235:genetics / Q000494:pharmacology; D040201:Platelet Membrane Glycoprotein IIb / Q000235:genetics / Q000378:metabolism; D020871:RNA Stability / Q000187:drug effects / Q000235:genetics; D015027:Zebrafish / Q000235:genetics / Q000378:metabolism; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D060172:Morpholinos; D040201:Platelet Membrane Glycoprotein IIb; D029961:Zebrafish Proteins
keywords: Antisense; Hemostasis; Thrombocytes; Vivo-morpholino; Zebrafish; αIIb