PUBLICATION
A Novel Zebrafish Model for Assessing In Vivo Delivery of Morpholino Oligomers
- Authors
- Kim, J., Clark, K., Barton, C., Tanguay, R., Moulton, H.
- ID
- ZDB-PUB-180902-6
- Date
- 2018
- Source
- Methods in molecular biology (Clifton, N.J.) 1828: 293-306 (Chapter)
- Registered Authors
- Clark, Karl, Tanguay, Robyn L.
- Keywords
- In vivo delivery, Morpholino oligomers, PPMO, Zebrafish model
- MeSH Terms
-
- Animals
- Exons
- Gene Expression
- Gene Transfer Techniques*
- Genes, Reporter
- Models, Animal
- Morpholinos/administration & dosage
- Morpholinos/genetics*
- Polymerase Chain Reaction
- Zebrafish/genetics*
- PubMed
- 30171549 Full text @ Meth. Mol. Biol.
Citation
Kim, J., Clark, K., Barton, C., Tanguay, R., Moulton, H. (2018) A Novel Zebrafish Model for Assessing In Vivo Delivery of Morpholino Oligomers. Methods in molecular biology (Clifton, N.J.). 1828:293-306.
Abstract
Morpholino oligomers have great therapeutic potential for treatment of a broad range of human diseases, including viral, bacterial, age-related, and genetic diseases, but they suffer from poor systemic delivery into cells. Although various approaches have been undertaken to address the delivery problem, it remains as the major barrier of morpholinos to be used as effective therapeutics. This slow development is in part due to the cost of materials and the animal models used for screening the efficacy and safety of those delivery approaches. The need to have an inexpensive vertebrate model for assessing in vivo delivery of morpholinos is evident. Therefore, we have produced a novel transgenic zebrafish model containing a dual reporter cassette for determination of in vivo delivery, bio-distribution, and safety of a morpholino. The levels of morpholino delivered to the cells in various tissues can be determined by changes in reporter gene expressions caused by morpholino-induced exon skipping. This chapter provides a description of the reagents, equipment, and procedure for successful retro-orbital injection of a peptide-conjugated morpholino into the blood stream of the adult zebrafish to cause targeted exon skipping in the heart of the zebrafish.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping