PUBLICATION

Regulating Gene Expression in Zebrafish Embryos Using Light-Activated, Negatively Charged Peptide Nucleic Acids

Authors
Tang, X., Maegawa, S., Weinberg, E.S., and Dmochowski, I.J.
ID
ZDB-PUB-070827-18
Date
2007
Source
Journal of the American Chemical Society   129(36): 11000-11001 (Journal)
Registered Authors
Maegawa, Shingo, Weinberg, Eric
Keywords
none
MeSH Terms
  • Animals
  • Gene Expression Regulation, Developmental/drug effects*
  • Glycoproteins/genetics*
  • Glycoproteins/metabolism
  • Intercellular Signaling Peptides and Proteins/genetics*
  • Intercellular Signaling Peptides and Proteins/metabolism
  • Light
  • Peptide Nucleic Acids/chemistry*
  • Peptide Nucleic Acids/pharmacology*
  • Peptide Nucleic Acids/radiation effects
  • RNA, Messenger
  • Zebrafish/embryology*
PubMed
17711280 Full text @ J. Am. Chem. Soc.
Abstract
We report the synthesis, characterization, and in vivo application of light-activated, negatively charged peptide nucleic acids (caged ncPNAs) for down-regulating gene expression during zebrafish embryonic development. The 18-mer ncPNAs were attached to a short complementary 2'-OMe RNA sense strand (sRNA) via a heterobifunctional photocleavable linker (PL). Caged ncPNAs were designed to target chordin and bozozok mRNA. Modest UV irradiation cleaved the photoactive linker, which yielded the less stable ncPNA/sRNA duplex (DeltaTm approximately -20 degrees C for chordin and -41 degrees C for bozozok). These caged ncPNAs were microinjected into zebrafish (Danio rerio) embryos at the one- or two-cell stage. The conjugate was nontoxic, and most embryos showed no phenotypic response at 24 h post fertilization (24 hpf), which indicated low background activity. Embryos that were photoactivated at 2 or 3 hpf developed phenotypic abnormalities typical of embryos with loss of function of the chordin or bozozok gene. Phenotypic results were further confirmed by in situ hybridization experiments using molecular markers. Caged ncPNAs provide a promising "on --> off" switch for down-regulating gene expression with high spatiotemporal control in biological systems.
Genes / Markers
Figures
Expression
Phenotype
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
Errata and Notes