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RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...

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Updated: Apr 1, 2026

Microinjection of mRNA and Morpholino Antisense Oligonucleotides in Zebrafish Embryos.

Microinjection of mRNA and Morpholino Antisense Oligonucleotides in Zebrafish Embryos.

Published on: May 7, 2009

Antisense Oligonucleotide-Mediated Transcript Knockdown in Zebrafish.

Andrea Pauli¹, Tessa G Montague¹, Kim A Lennox²

¹Department of Molecular and Cellular Biology, Harvard University, Cambridge, Masschusetts, United States of America.

|October 6, 2015

Summary

This summary is machine-generated.

Antisense oligonucleotides (ASOs) effectively knocked down developmental genes in zebrafish, mirroring known loss-of-function phenotypes. This RNA-targeting method shows promise for gene knockdown studies in zebrafish, including long noncoding RNAs.

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Micromanipulation of Gene Expression in the Adult Zebrafish Brain Using Cerebroventricular Microinjection of Morpholino Oligonucleotides

Micromanipulation of Gene Expression in the Adult Zebrafish Brain Using Cerebroventricular Microinjection of Morpholino Oligonucleotides

Published on: May 23, 2013

Area of Science:

Molecular Biology
Developmental Biology
Zebrafish Genetics

Background:

Antisense oligonucleotides (ASOs) are RNA-DNA hybrids inducing RNA degradation via RNase H.
ASOs are established gene knockdown tools in various model systems.

Purpose of the Study:

To evaluate the efficacy of ASOs as gene knockdown reagents in zebrafish.
To compare ASO-induced phenotypes with existing mutant and morpholino data.

Main Methods:

ASOs targeting 20 developmental genes were administered to zebrafish embryos.
Morphological changes were documented and compared to known phenotypes.
Knockdown efficiency and toxicity were assessed at various concentrations.

Main Results:

ASO treatment reproduced loss-of-function phenotypes for several genes (oep, chordin, dnd, ctnnb2, bmp7a, alk8, smad2, smad5) in a dose-dependent manner.
ASOs successfully knocked down both maternal/zygotic transcripts and the MALAT1 lncRNA.
ASO efficacy was observed within a narrow concentration range, with toxicity at higher doses.

Conclusions:

ASOs are effective for gene knockdown in zebrafish, recapitulating known developmental phenotypes.
ASOs offer a valuable tool for studying gene function, including lncRNAs, in zebrafish models.
Careful optimization of ASO concentration is necessary to balance efficacy and toxicity.