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PLoS One. 2015 Oct 5;10(10):e0139504. doi: 10.1371/journal.pone.0139504. eCollection 2015.

Antisense Oligonucleotide-Mediated Transcript Knockdown in Zebrafish.

Author information

1
Department of Molecular and Cellular Biology, Harvard University, Cambridge, Masschusetts, United States of America.
2
Integrated DNA Technologies, Inc., Coralville, Iowa, United States of America.

Abstract

Antisense oligonucleotides (ASOs) are synthetic, single-strand RNA-DNA hybrids that induce catalytic degradation of complementary cellular RNAs via RNase H. ASOs are widely used as gene knockdown reagents in tissue culture and in Xenopus and mouse model systems. To test their effectiveness in zebrafish, we targeted 20 developmental genes and compared the morphological changes with mutant and morpholino (MO)-induced phenotypes. ASO-mediated transcript knockdown reproduced the published loss-of-function phenotypes for oep, chordin, dnd, ctnnb2, bmp7a, alk8, smad2 and smad5 in a dosage-sensitive manner. ASOs knocked down both maternal and zygotic transcripts, as well as the long noncoding RNA (lncRNA) MALAT1. ASOs were only effective within a narrow concentration range and were toxic at higher concentrations. Despite this drawback, quantitation of knockdown efficiency and the ability to degrade lncRNAs make ASOs a useful knockdown reagent in zebrafish.

PMID:
26436892
PMCID:
PMC4593562
DOI:
10.1371/journal.pone.0139504
[Indexed for MEDLINE]
Free PMC Article

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