Quantitating plant microRNA-mediated target repression using a dual-luciferase transient expression system

Qikun Liu; Michael J. Axtell

doi:10.1007/978-1-4939-2444-8_14

Quantitating plant microRNA-mediated target repression using a dual-luciferase transient expression system

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Abstract

microRNA (miRNA) mediated repression of target genes plays essential roles in a variety of functions in plants. An easy-to-use method that can effectively validate functional miRNA–target interactions in plants thus is of particular interest. Here, we describe an Agrobacterium tumefaciens -mediated in vivo assay utilizing a dual-luciferase reporter system. With this method, the strength of miRNA-mediated target repression can be quantifi ed at both the mRNA (via qRT-PCR) and protein (via dual-luciferase assay) levels quickly and accurately.

Original language	English (US)
Pages (from-to)	287-303
Number of pages	17
Journal	Methods in Molecular Biology
Volume	1284
DOIs	https://doi.org/10.1007/978-1-4939-2444-8_14
State	Published - Jan 1 2015

All Science Journal Classification (ASJC) codes

Molecular Biology
Genetics

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10.1007/978-1-4939-2444-8_14

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abstract = "microRNA (miRNA) mediated repression of target genes plays essential roles in a variety of functions in plants. An easy-to-use method that can effectively validate functional miRNA–target interactions in plants thus is of particular interest. Here, we describe an Agrobacterium tumefaciens -mediated in vivo assay utilizing a dual-luciferase reporter system. With this method, the strength of miRNA-mediated target repression can be quantifi ed at both the mRNA (via qRT-PCR) and protein (via dual-luciferase assay) levels quickly and accurately.",

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AB - microRNA (miRNA) mediated repression of target genes plays essential roles in a variety of functions in plants. An easy-to-use method that can effectively validate functional miRNA–target interactions in plants thus is of particular interest. Here, we describe an Agrobacterium tumefaciens -mediated in vivo assay utilizing a dual-luciferase reporter system. With this method, the strength of miRNA-mediated target repression can be quantifi ed at both the mRNA (via qRT-PCR) and protein (via dual-luciferase assay) levels quickly and accurately.

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Quantitating plant microRNA-mediated target repression using a dual-luciferase transient expression system

Abstract

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