Detection of the argonaute protein Ago2 and microRNAs in the RNA induced silencing complex (RISC) using a monoclonal antibody

Authors

K. Ikeda
M. Satoh
Kaleb M. Pauley, Cedarville UniversityFollow
M. J. Fritzler
W. H. Reeves
E. K. Chan

Document Type

Article

Publication Date

10-4-2006

Journal Title

Journal of Immunological Methods

Volume

317

Issue

1-2

First Page

38

Last Page

44

Abstract

MicroRNAs (miRNAs) are short RNA molecules responsible for post-transcriptional gene silencing by the degradation or translational inhibition of their target messenger RNAs (mRNAs). This process of gene silencing, known as RNA interference (RNAi), is mediated by highly conserved Argonaute (Ago) proteins which are the key components of the RNA induced silencing complex (RISC). In humans, Ago2 is responsible for the endonuclease cleavage of targeted mRNA and it interacts with the mRNAbinding protein GW182, which is a marker for cytoplasmic foci referred to as GW bodies (GWBs). We demonstrated that the antiAgo2 monoclonal antibody 4F9 recognized GWBs in a cell cycle dependent manner and was capable of capturing miRNAs associated with Ago2. Since Ago2 protein is the effector protein of RNAi, anti-Ago2 monoclonal antibody may be useful in capturing functional miRNAs.

Keywords

Argonaut protein, GW182, GW bodies, Monoclonal antibody, MicroRNA, P body, RNA interference

Recommended Citation

Ikeda, K.; Satoh, M.; Pauley, Kaleb M.; Fritzler, M. J.; Reeves, W. H.; and Chan, E. K., "Detection of the argonaute protein Ago2 and microRNAs in the RNA induced silencing complex (RISC) using a monoclonal antibody" (2006). Science and Mathematics Faculty Publications. 197.
https://digitalcommons.cedarville.edu/science_and_mathematics_publications/197