Detection of slicer activity by immunopurified plant ARGONAUTE1

Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review

Standard

Detection of slicer activity by immunopurified plant ARGONAUTE1. / Arribas Hernandez, Laura; Vigh, Maria Louisa; Brodersen, Peter.

Plant MicroRNAs: Methods and Protocols. ed. / Stefan de Folter. Humana Press, 2019. p. 295-316 (Methods in Molecular Biology, Vol. 1932).

Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review

Harvard

Arribas Hernandez, L, Vigh, ML & Brodersen, P 2019, Detection of slicer activity by immunopurified plant ARGONAUTE1. in SD Folter (ed.), Plant MicroRNAs: Methods and Protocols. Humana Press, Methods in Molecular Biology, vol. 1932, pp. 295-316. https://doi.org/10.1007/978-1-4939-9042-9_22

APA

Arribas Hernandez, L., Vigh, M. L., & Brodersen, P. (2019). Detection of slicer activity by immunopurified plant ARGONAUTE1. In S. D. Folter (Ed.), Plant MicroRNAs: Methods and Protocols (pp. 295-316). Humana Press. Methods in Molecular Biology Vol. 1932 https://doi.org/10.1007/978-1-4939-9042-9_22

Vancouver

Arribas Hernandez L, Vigh ML, Brodersen P. Detection of slicer activity by immunopurified plant ARGONAUTE1. In Folter SD, editor, Plant MicroRNAs: Methods and Protocols. Humana Press. 2019. p. 295-316. (Methods in Molecular Biology, Vol. 1932). https://doi.org/10.1007/978-1-4939-9042-9_22

Author

Arribas Hernandez, Laura ; Vigh, Maria Louisa ; Brodersen, Peter. / Detection of slicer activity by immunopurified plant ARGONAUTE1. Plant MicroRNAs: Methods and Protocols. editor / Stefan de Folter. Humana Press, 2019. pp. 295-316 (Methods in Molecular Biology, Vol. 1932).

Bibtex

@inbook{a462550e85c048d1b62ea77c125b1356,
title = "Detection of slicer activity by immunopurified plant ARGONAUTE1",
abstract = "Small RNA-guided endonucleolysis (“slicing”) of target mRNA is the signature biochemical activity underlying many RNA silencing phenomena. The catalytic slicer activity resides in Argonaute (AGO) proteins. Here, we present two protocols to detect microRNA-guided slicer activity of AGO1 immunopurified from Arabidopsis tissues. The first uses radioactive, cap-labeled RNA substrates produced by in vitro transcription of RNA fragments corresponding to endogenous target sites flanked by 100–200 nucleotides of target sequence. The second protocol uses similarly designed but shorter (around 50 nt) fluorescently labeled RNA. Advantages and disadvantages of the two setups are also discussed.",
keywords = "Arabidopsis, Argonaute, Endonucleolysis, In vitro assay, miRNA, Slicing",
author = "{Arribas Hernandez}, Laura and Vigh, {Maria Louisa} and Peter Brodersen",
year = "2019",
doi = "10.1007/978-1-4939-9042-9_22",
language = "English",
isbn = "978-1-4939-9041-2",
series = "Methods in Molecular Biology",
publisher = "Humana Press",
pages = "295--316",
editor = "Folter, {Stefan de}",
booktitle = "Plant MicroRNAs",
address = "United States",

}

RIS

TY - CHAP

T1 - Detection of slicer activity by immunopurified plant ARGONAUTE1

AU - Arribas Hernandez, Laura

AU - Vigh, Maria Louisa

AU - Brodersen, Peter

PY - 2019

Y1 - 2019

N2 - Small RNA-guided endonucleolysis (“slicing”) of target mRNA is the signature biochemical activity underlying many RNA silencing phenomena. The catalytic slicer activity resides in Argonaute (AGO) proteins. Here, we present two protocols to detect microRNA-guided slicer activity of AGO1 immunopurified from Arabidopsis tissues. The first uses radioactive, cap-labeled RNA substrates produced by in vitro transcription of RNA fragments corresponding to endogenous target sites flanked by 100–200 nucleotides of target sequence. The second protocol uses similarly designed but shorter (around 50 nt) fluorescently labeled RNA. Advantages and disadvantages of the two setups are also discussed.

AB - Small RNA-guided endonucleolysis (“slicing”) of target mRNA is the signature biochemical activity underlying many RNA silencing phenomena. The catalytic slicer activity resides in Argonaute (AGO) proteins. Here, we present two protocols to detect microRNA-guided slicer activity of AGO1 immunopurified from Arabidopsis tissues. The first uses radioactive, cap-labeled RNA substrates produced by in vitro transcription of RNA fragments corresponding to endogenous target sites flanked by 100–200 nucleotides of target sequence. The second protocol uses similarly designed but shorter (around 50 nt) fluorescently labeled RNA. Advantages and disadvantages of the two setups are also discussed.

KW - Arabidopsis

KW - Argonaute

KW - Endonucleolysis

KW - In vitro assay

KW - miRNA

KW - Slicing

U2 - 10.1007/978-1-4939-9042-9_22

DO - 10.1007/978-1-4939-9042-9_22

M3 - Book chapter

C2 - 30701509

AN - SCOPUS:85060922812

SN - 978-1-4939-9041-2

T3 - Methods in Molecular Biology

SP - 295

EP - 316

BT - Plant MicroRNAs

A2 - Folter, Stefan de

PB - Humana Press

ER -

ID: 214511805