PUBLICATION

Efficient Transfection Strategy for the Spatiotemporal Control of Gene Expression in Zebrafish

Authors
Ando, H., and Okamoto, H.
ID
ZDB-PUB-060419-5
Date
2006
Source
Marine biotechnology (New York, N.Y.)   8(3): 295-303 (Journal)
Registered Authors
Ando, Hideki
Keywords
Gene expression, lipofection, morpholino oligonucleotides, transfection, zebrafish
MeSH Terms
  • Animals
  • Brain/embryology
  • Brain/metabolism
  • DNA/genetics
  • DNA/metabolism
  • Eye/embryology
  • Eye/metabolism
  • Gene Expression Regulation, Developmental*
  • Larva/genetics
  • Larva/metabolism
  • Oligonucleotides, Antisense/genetics
  • Oligonucleotides, Antisense/metabolism
  • RNA, Messenger/genetics
  • RNA, Messenger/metabolism
  • Transfection/methods*
  • Zebrafish/embryology
  • Zebrafish/genetics*
PubMed
16614871 Full text @ Mar. Biotechnol.
Abstract
Functional analyses of gene function by knockdown and expression approaches strongly enhance the genetic study of development. In vivo application of the introduction of inhibitors of gene expression, mRNA, and expression constructs in the target region make it possible to perform region- and stage-specific regulation of gene function in a simple manner. As a basic tool for the conditional regulation of gene expression in target tissue, we present methods for the efficient introduction of antisense morpholino oligonucleotide (MO), mRNA, and expression plasmid constructs into early and later stage zebrafish embryo and larva. Lipofection of a neuron-specific expression construct plasmid encoding green fluorescent protein (GFP) into optic vesicle resulted in clear GFP expression in the retinotectal pathway in hatched larva. Co-lipofection of MO and GFP mRNA to the presumptive head region resulted in brain-specific knockdown of the gene in mid-stage embryos.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping