Department of Chemistry & Biochemistry Seminar
Wednesday, September 24, 2014
Detection of Argonaute (Ago) Protein Associated MicroRNA by Combining Anti-Ago
Antibody Recognition with Real-Time RT-PCR
Brian Coley
MS Student
UNCG
MicroRNAs (miRNAs) are small noncoding RNA that range between ~19-25 nucleotides
in length. In recent years, scientists have observed that these small RNA molecules
exist in the extracellular environment within eukaryotic organisms. Furthermore, these
microRNA molecules are known to associate with a family of proteins named Argonaute
(Ago) proteins. These microRNA/Ago protein complexes are the core of a larger
assembly of proteins that compose the RNA Induced Silencing Complex (RISC). The
RISC has exhibited the ability to inhibit the translation of, or cleave, its target messenger
RNA (mRNA), the latter of which being exclusive only to Ago2. It has also been
observed in the literature that these miRNA/Ago complexes often target genomic
regions associated with various cancers in humans and, currently, more than 2000
miRNAs have been discovered and published in the literature.
Current methods for analyzing miRNA expression involve total RNA extraction using
methods such as ethanol precipitation. However, total RNA extraction does not take into
consideration that the functional component of post-transcriptional inhibition is indeed that
miRNA/Ago complex and not the miRNA alone. In this study, we investigate a novel
method to capture the active miRNA/Ago2 complex and quantitate the associated
miRNAs by utilizing an antibody against Ago2 and subsequent application of real-time
PCR.
1:00 p.m. Sullivan Science Building * Wachovia Lecture Hall (Room 201)