PUBLICATION
Disease Modeling by Gene Targeting Using MicroRNAs
- Authors
- Lan, C.C., Leong, I.U., Lai, D., and Love, D.R.
- ID
- ZDB-PUB-111012-22
- Date
- 2011
- Source
- Methods in cell biology 105: 419-436 (Chapter)
- Registered Authors
- Love, Donald R.
- Keywords
- cleavage, intron, microinjection, silencing, transcriptional, vector
- MeSH Terms
-
- Animals
- Disease Models, Animal
- Embryo, Nonmammalian
- Gene Targeting/methods*
- Genes, Reporter
- Genetic Diseases, Inborn/genetics*
- Genetic Diseases, Inborn/pathology
- Haploinsufficiency
- Humans
- MicroRNAs*/genetics
- MicroRNAs*/metabolism
- Microinjections/methods*
- Microscopy, Fluorescence
- Molecular Biology/methods*
- Molecular Imaging/methods*
- Oligodeoxyribonucleotides/genetics
- Oligodeoxyribonucleotides/metabolism
- Plasmids
- RNA Interference*
- RNA, Messenger/genetics
- RNA, Messenger/metabolism
- RNA, Small Interfering*/genetics
- RNA, Small Interfering*/metabolism
- Ribonuclease III/genetics
- Ribonuclease III/metabolism
- Zebrafish/embryology
- Zebrafish/genetics*
- Zebrafish/metabolism
- PubMed
- 21951541 Full text @ Meth. Cell. Biol.
Citation
Lan, C.C., Leong, I.U., Lai, D., and Love, D.R. (2011) Disease Modeling by Gene Targeting Using MicroRNAs. Methods in cell biology. 105:419-436.
Abstract
Zebrafish have proved to be a popular species for the modeling of human disease. In this context, there is a need to move beyond chemical-based mutagenesis and develop tools that target genes that are orthologous to those that are implicated in human heritable diseases. Targeting can take the form of creating mutations that are nonsense or mis-sense, or to mimic haploinsufficiency through the regulated expression of RNA effector molecules. In terms of the latter, we describe here the development and investigation of microRNA (miRNA)-based directed gene silencing methods in zebrafish. Unlike small interfering RNAs (siRNAs), miRNA-based methods offer temporal and spatial regulation of gene silencing. Proof-of-concept experiments demonstrate the efficacy of the method in zebrafish embryos, which provide the foundation for developing disease models using miRNA-based gene-targeting.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping