Poster abstracts
Poster number 59 submitted by Hannah Hayth
Bioconjugation of functionalized oligodeoxynucleotides to fluorescence reporters
Hannah L. Hayth (Ball State University), Erwin Doe (Ball State University), Emil Khisamutdinov (Ball State University)
Abstract:
In the field of oligonucleotide (ODN) therapeutics, there is an imperative need to improve the ODNs properties by either chemical modification of the oligonucleotides structure or covalently linking reporter or therapeutic moieties with biologically relevant properties. Chemical conjugation can thus, significantly improve the intrinsic property not only of ODNs but also reporter/therapeutic molecules. Bioconjugation of nucleic acids to small molecules also serve as nano-delivery facilities to transport drugs to specific targets. This study deployed azide–alkyne cycloaddition, a click reaction, to successfully conjugate a Cyanine-3 alkyne moiety to an azide-functionalized ODN 12-mer, as well as 3–azido–7–hydroxy–chromen–2–one, a coumarin azide to an alkyne functionalized ODN 12-mer. The clicked products were subsequently hybridized with DNA nanoparticles to serves as fluorphore-labeled drug delivery units. The application of Huisgen 1,3–dipolar cycloaddition (HDC) of azides to terminal alkynes via a Cu(I)–catalyzed reaction pathway has been scientifically established to produce high yield and stable 1,4–disubstituted 1,2,3–triazoles. Post-conjugation investigation of the clicked products via fluorescence spectroscopy and denaturing polyacrylamide gel electrophoresis demonstrated a drastic amplitude of fluorescent intensity by multiple folds of the reporter molecules.
Keywords: bioconjugation, click chemistry, nucleic acid nanoparticles