In Vivo Testing of MicroRNA-Mediated Gene Knockdown in Zebrafish

The zebrafish (Danio rerio) has become an attractive model for human disease modeling as there are a large number of orthologous genes that encode similar proteins to those found in humans. The number of tools available to manipulate the zebrafish genome is limited and many currently used techniques...

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Bibliographic Details
Published inJournal of biomedicine & biotechnology Vol. 2012; no. 2012; pp. 1 - 7
Main Authors Leong, Ivone Un San, Lan, Chuan-Ching, Skinner, Jonathan R., Shelling, Andrew N., Love, Donald R.
Format Journal Article
LanguageEnglish
Published Cairo, Egypt Hindawi Puplishing Corporation 01.01.2012
Hindawi Publishing Corporation
Subjects
Analysis of Variance
Animals
Disease Models, Animal
Embryo, Nonmammalian
Ether-A-Go-Go Potassium Channels - genetics
Ether-A-Go-Go Potassium Channels - metabolism
Gene Knockdown Techniques - methods
Green Fluorescent Proteins - genetics
Green Fluorescent Proteins - metabolism
Humans
Long QT Syndrome
Mice
MicroRNAs - chemistry
MicroRNAs - genetics
RNA Interference
Zebrafish
Zebrafish Proteins - genetics
Zebrafish Proteins - metabolism
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Summary:The zebrafish (Danio rerio) has become an attractive model for human disease modeling as there are a large number of orthologous genes that encode similar proteins to those found in humans. The number of tools available to manipulate the zebrafish genome is limited and many currently used techniques are only effective during early development (such as morpholino-based antisense technology) or it is phenotypically driven and does not offer targeted gene knockdown (such as chemical mutagenesis). The use of RNA interference has been met with controversy as off-target effects can make interpreting phenotypic outcomes difficult; however, this has been resolved by creating zebrafish lines that contain stably integrated miRNA constructs that target the desired gene of interest. In this study, we show that a commercially available miRNA vector system with a mouse-derived miRNA backbone is functional in zebrafish and is effective in causing eGFP knockdown in a transient in vivo eGFP sensor assay system. We chose to apply this system to the knockdown of transcripts that are implicated in the human cardiac disorder, Long QT syndrome.
Bibliography:Academic Editor: Kurt Bürki
ISSN:1110-7243
1110-7251
DOI:10.1155/2012/350352