NDLI: Testing the efficiency of dsRNAi constructs in vivo: A transient expression assay based on two fluorescent proteins

Content Provider	Springer Nature Link
Author	Panstruga, Ralph Chul Kim, Min Cho, Moo Je Schulze Lefert, Paul
Copyright Year	2003
Abstract	Double-stranded RNA interference technology has recently been shown to be a powerful tool to silence gene expression in various organisms, including plants. Sustained double-stranded RNA interference mediated gene silencing is normally triggered by hairpin RNAs generated by in vivo transcription of inverted repeat DNA constructs. To test the efficiency of inverted repeat constructs for their in vivo gene silencing capability, we developed an assay that is based on transient expression in single cells using two fluorescent reporter proteins with different emission spectra. We co-expressed one fluorescent protein as marker for transfected cells and the second as translational fusion with the target gene. Co-transfection of a vector mediating expression of inverted repeats of the target gene resulted in a specific decrease or undetectable fusion protein fluorescence in the majority of transfected cells.
Starting Page	135
Ending Page	140
Page Count	6
File Format	PDF
ISSN	03014851
Journal	Molecular Biology Reports
Volume Number	30
Issue Number	3
e-ISSN	15734978
Language	English
Publisher	Kluwer Academic Publishers
Publisher Date	2003-01-01
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Animal Biochemistry Animal Anatomy / Morphology / Histology
Content Type	Text
Resource Type	Article
Subject	Genetics Medicine Molecular Biology

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Testing the efficiency of dsRNAi constructs in vivo: A transient expression assay based on two fluorescent proteins