Antisense Oligonucleotide-Mediated Transcript Knockdown in Zebrafish
JournalArticle (Originalarbeit in einer wissenschaftlichen Zeitschrift)
 
ID 4519618
Author(s) Pauli, Andrea; Montague, Tessa G.; Lennox, Kim A.; Behlke, Mark A.; Schier, Alexander F.
Author(s) at UniBasel Schier, Alexander
Year 2015
Title Antisense Oligonucleotide-Mediated Transcript Knockdown in Zebrafish
Journal PloS one
Volume 10
Number 10
Pages / Article-Number e0139504
Mesh terms Animals; Embryonic Development, genetics; Feasibility Studies; Female; Gene Knockdown Techniques; Male; Morpholinos, genetics, pharmacology; Oligonucleotides, Antisense, genetics, pharmacology; RNA, Long Noncoding, genetics; RNA, Messenger, antagonists & inhibitors, genetics; Transcription, Genetic; Zebrafish, embryology, genetics; Zebrafish Proteins, genetics; Zygote
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.
Publisher PUBLIC LIBRARY SCIENCE
ISSN/ISBN 1932-6203
URL https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0139504
edoc-URL https://edoc.unibas.ch/74125/
Full Text on edoc No
Digital Object Identifier DOI 10.1371/journal.pone.0139504
PubMed ID http://www.ncbi.nlm.nih.gov/pubmed/26436892
ISI-Number WOS:000362499200030
Document type (ISI) Journal Article
 
   

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