Date of Award

1-1-2012

Language

English

Document Type

Master's Thesis

Degree Name

Master of Science (MS)

College/School/Department

Department of Chemistry

Content Description

1 online resource (vi, 44 pages) : illustrations (some color)

Dissertation/Thesis Chair

Li Niu

Committee Members

Li Niu, Jayanti Pande

Keywords

AMPA receptor, flop, Glutamate receptor, RNA Aptamer, SELEX, Glutamic acid, Neurotransmitter receptors, Oligonucleotides

Subject Categories

Biochemistry | Chemistry

Abstract

The á-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors play an important role in neurotransmission and synaptic plasticity in the central nervous system. Excessive activity of these receptors has been implicated in some neurodegenerative diseases, such as Parkinson's disease, ischemic stroke and amyotrophic lateral sclerosis (ALS). AMPA receptor inhibitors are therefore drug candidates for potential treatment of these neurological disorders and diseases. The objective of my MS thesis work is to develop a stable aptamer so that it can be tested and used in vivo where the aptamer must be exposed to ribonucleases. To do this, I prepared a library containing RNAs with 2'-Fluoro and 2'-O-Methoxy nucleotides. By re-running an in vitro selection method called systematic evolution of ligands by exponential enrichment (SELEX) with this chemically modified RNA library, I have identified two chemically modified RNA aptamers with consensus sequences against the GluA2Qflop receptor. The flop isoform of GluA2Q or GluA2Qflop AMPA receptor subunit was chosen because GluA2 is a key AMPA receptor subunit mediating excitotoxicity. In a preliminary test, I found that one RNA aptamer is stable in cerebrospinal fluid with a half-life of >3 days. In contrast, unmodified RNA is degraded in a few minutes. Furthermore, this RNA is indeed inhibitory to the GluA2 receptor, the selection target. The significance of my research is that once fully developed, a chemically modified, stable aptamer targeting AMPA receptors is a real drug candidate amenable to be tested under in vivo conditions, such as in cellular and animal models and ultimately in human trials as drug.

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