Title of Abstract

Using Nucleic Acid-Gold Nanoparticle Conjugates in the Fight Against Bacteria that are Resistant to Tetracycline Antibiotics

Faculty Mentor

One WU mentor: Timea Fernandez, Ph.D.; fernandezt@winthrop.edu

College

College of Arts and Sciences

Department

Chemistry, Physics and Geology

Faculty Mentor

Timea Fernandez, Ph.D.

Abstract

Antibiotic resistance is major problem in modern medicine. Seventy percent of bacterial strains are resistant to at least one antibiotic, making treatment of bacterial infections ever more expensive and difficult. Currently, we are investigating the therapeutic potency of nucleic acid-gold nanoparticle conjugates as treatments against bacteria that are resistant to the antibiotic tetracycline. We hypothesize that by attaching RNA that binds to tetracycline to silver or gold nanoparticles the resulting conjugates will work as a “Trojan-horse” tetracycline-delivery vehicle that smuggles the antibiotic into the cell without being detected by cellular defense systems. Moreover, we reason that silver or gold ions released by the nanoparticles add to the antimicrobial effects of tetracycline. To demonstrate the viability of this idea, we set out to generate a nuclease resistant variant of the tetracycline binding RNA ykkCD and attach it to gold nanoparticles. During the summer we furthered the progress of this research in three major ways. First, we optimized conditions used for the polymerase chain reaction that generates the DNA template for RNA synthesis. Second, we found the most efficient way to synthesize RNA containing modified nucleotides. Usage of modified nucleotides is necessary to prevent degradation of the tetracycline-binding RNA by cellular nucleases. Third, we optimized a stability assay to verify that the modified RNAs are stable in cellular environments. Fourth, we attached the tetracycline binding RNA to gold nanoparticles. The next steps in the process would be to test the potency of this modified RNA-gold nanoparticle conjugate against a tetracycline-resistant strain of the bacteria E. coli.

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Grant Support

NIH-NCBI, SC EPSCoR, and INBRE

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Using Nucleic Acid-Gold Nanoparticle Conjugates in the Fight Against Bacteria that are Resistant to Tetracycline Antibiotics

Antibiotic resistance is major problem in modern medicine. Seventy percent of bacterial strains are resistant to at least one antibiotic, making treatment of bacterial infections ever more expensive and difficult. Currently, we are investigating the therapeutic potency of nucleic acid-gold nanoparticle conjugates as treatments against bacteria that are resistant to the antibiotic tetracycline. We hypothesize that by attaching RNA that binds to tetracycline to silver or gold nanoparticles the resulting conjugates will work as a “Trojan-horse” tetracycline-delivery vehicle that smuggles the antibiotic into the cell without being detected by cellular defense systems. Moreover, we reason that silver or gold ions released by the nanoparticles add to the antimicrobial effects of tetracycline. To demonstrate the viability of this idea, we set out to generate a nuclease resistant variant of the tetracycline binding RNA ykkCD and attach it to gold nanoparticles. During the summer we furthered the progress of this research in three major ways. First, we optimized conditions used for the polymerase chain reaction that generates the DNA template for RNA synthesis. Second, we found the most efficient way to synthesize RNA containing modified nucleotides. Usage of modified nucleotides is necessary to prevent degradation of the tetracycline-binding RNA by cellular nucleases. Third, we optimized a stability assay to verify that the modified RNAs are stable in cellular environments. Fourth, we attached the tetracycline binding RNA to gold nanoparticles. The next steps in the process would be to test the potency of this modified RNA-gold nanoparticle conjugate against a tetracycline-resistant strain of the bacteria E. coli.