PUBLICATION
Knockdown of αIIb by RNA degradation by delivering deoxyoligonucleotides piggybacked with control vivo-morpholinos into zebrafish thrombocytes
- Authors
- Sundaramoorthi, H., Khandekar, G., Kim, S., Jagadeeswaran, P.
- ID
- ZDB-PUB-140820-7
- Date
- 2015
- Source
- Blood cells, molecules & diseases 54(1): 78-83 (Journal)
- Registered Authors
- Jagadeeswaran, Pudur, Khandekar, Gauri, Kim, Seongcheol
- Keywords
- Antisense, Hemostasis, Thrombocytes, Vivo-morpholino, Zebrafish, αIIb
- MeSH Terms
-
- Animals
- Blood Platelets/metabolism*
- Gene Knockdown Techniques/methods*
- Morpholinos*/genetics
- Morpholinos*/pharmacology
- Platelet Membrane Glycoprotein IIb*/genetics
- Platelet Membrane Glycoprotein IIb*/metabolism
- RNA Stability*/drug effects
- RNA Stability*/genetics
- Zebrafish*/genetics
- Zebrafish*/metabolism
- Zebrafish Proteins*/genetics
- Zebrafish Proteins*/metabolism
- PubMed
- 25135204 Full text @ Blood Cells Mol. Dis.
Citation
Sundaramoorthi, H., Khandekar, G., Kim, S., Jagadeeswaran, P. (2015) Knockdown of αIIb by RNA degradation by delivering deoxyoligonucleotides piggybacked with control vivo-morpholinos into zebrafish thrombocytes. Blood cells, molecules & diseases. 54(1):78-83.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping