Knockdown of αIIb by RNA degradation by delivering deoxyoligonucleotides piggybacked with control vivo-morpholinos into zebrafish thrombocytes

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 kn...

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Bibliographic Details
Published in:Blood cells, molecules, & diseases molecules, & diseases, 2015-01, Vol.54 (1), p.78-83
Main Authors: Sundaramoorthi, Hemalatha, Khandekar, Gauri, Kim, Seongcheol, Jagadeeswaran, Pudur
Format: Article
Language:English
Subjects:
Animals
Antisense
Blood Platelets - metabolism
Gene Knockdown Techniques - methods
Hemostasis
Morpholinos - genetics
Morpholinos - pharmacology
Platelet Membrane Glycoprotein IIb - genetics
Platelet Membrane Glycoprotein IIb - metabolism
RNA Stability - drug effects
RNA Stability - genetics
Thrombocytes
Vivo-morpholino
Zebrafish
Zebrafish - genetics
Zebrafish - metabolism
Zebrafish Proteins - genetics
Zebrafish Proteins - metabolism
αIIb
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Summary: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.
ISSN:1079-9796
1096-0961
DOI:10.1016/j.bcmd.2014.07.016