PUBLICATION
Microinjection of mRNA and morpholino antisense oligonucleotides in zebrafish embryos
- Authors
- Yuan, S., and Sun, Z.
- ID
- ZDB-PUB-090616-13
- Date
- 2009
- Source
- Journal of visualized experiments : JoVE (27): (Journal)
- Registered Authors
- Sun, Zhaoxia, Yuan, Shiaulou
- Keywords
- none
- MeSH Terms
-
- Animals
- Green Fluorescent Proteins/genetics
- Microinjections/methods*
- Microscopy, Fluorescence/methods
- Oligonucleotides, Antisense/administration & dosage*
- Oligonucleotides, Antisense/genetics
- RNA, Messenger/administration & dosage*
- RNA, Messenger/genetics
- Zebrafish/embryology*
- Zebrafish/genetics*
- PubMed
- 19488022 Full text @ J. Vis. Exp.
Citation
Yuan, S., and Sun, Z. (2009) Microinjection of mRNA and morpholino antisense oligonucleotides in zebrafish embryos. Journal of visualized experiments : JoVE. (27).
Abstract
An essential tool for investigating the role of a gene during development is the ability to perform gene knockdown, overexpression, and misexpression studies. In zebrafish (Danio rerio), microinjection of RNA, DNA, proteins, antisense oligonucleotides and other small molecules into the developing embryo provides researchers a quick and robust assay for exploring gene function in vivo. In this video-article, we will demonstrate how to prepare and microinject in vitro synthesized EGFP mRNA and a translational-blocking morpholino oligo against pkd2, a gene associated with autosomal dominant polycystic kidney disease (ADPKD), into 1-cell stage zebrafish embryos. We will then analyze the success of the mRNA and morpholino microinjections by verifying GFP expression and phenotype analysis. Broad applications of this technique include generating transgenic animals and germ-line chimeras, cell-fate mapping and gene screening. Herein we describe a protocol for overexpression of EGFP and knockdown of pkd2 by mRNA and morpholino oligonucleotide injection.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping