Title of Abstract

Using Tetracycline-Binding Nucleic Acid Aptamers as Trojan-Horse Tetracycline Delivery Vehicles in the Fight Against Drug-Resistant Bacteria

Poster Number

67

Session Title

Poster Session 2

Faculty Sponsor (for work done with a non-Winthrop mentor)

Timea Fernandez, Ph.D. & Victoria Frost, Ph.D.

College

College of Arts and Sciences

Department

Chemistry, Physics & Geology

Abstract

Illnesses caused by bacteria are now a major public health concern since microorganisms have become increasingly resistant to available antibiotics. As this has occurred big pharma has gradually shifted its focus from developing drugs that cure diseases to those that treat chronic conditions. Thus, rediscovering old drugs and using them for new purposes have become more important. The ultimate goal of this project is to use nucleic acid aptamer-nanoparticle conjugates as vehicles that deliver the antibiotics to cells that are resistant to them. We investigated the therapeutic potency of nucleic acid- silver/gold nanoparticle conjugates as treatments against bacteria that are resistant to the antibiotic tetracycline. We hypothesized that by attaching nucleic acid aptamers that bind 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. In addition we reasoned that the silver or gold ions released by the nanoparticles will add to the antimicrobial effects of tetracycline. To further prove the viability of this idea we tested three tetracycline binding nucleic acid aptamers. PCR conditions were optimized to make DNA template for RNA synthesis, transcription of RNA aptamers using modified nucleotides was performed, aptamer/linker annealing conditions were optimized, and finally we attached aptamers to gold/silver nanoparticles. In addition, we developed serum stability assays to demonstrate that the RNA aptamers are not degraded by cellular nucleases thus able exert their function in live cells. We are currently testing antimicrobial effect of aptamer nanoparticle conjugates using E. coli 5922.

Start Date

15-4-2022 12:00 PM

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Apr 15th, 12:00 PM

Using Tetracycline-Binding Nucleic Acid Aptamers as Trojan-Horse Tetracycline Delivery Vehicles in the Fight Against Drug-Resistant Bacteria

Illnesses caused by bacteria are now a major public health concern since microorganisms have become increasingly resistant to available antibiotics. As this has occurred big pharma has gradually shifted its focus from developing drugs that cure diseases to those that treat chronic conditions. Thus, rediscovering old drugs and using them for new purposes have become more important. The ultimate goal of this project is to use nucleic acid aptamer-nanoparticle conjugates as vehicles that deliver the antibiotics to cells that are resistant to them. We investigated the therapeutic potency of nucleic acid- silver/gold nanoparticle conjugates as treatments against bacteria that are resistant to the antibiotic tetracycline. We hypothesized that by attaching nucleic acid aptamers that bind 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. In addition we reasoned that the silver or gold ions released by the nanoparticles will add to the antimicrobial effects of tetracycline. To further prove the viability of this idea we tested three tetracycline binding nucleic acid aptamers. PCR conditions were optimized to make DNA template for RNA synthesis, transcription of RNA aptamers using modified nucleotides was performed, aptamer/linker annealing conditions were optimized, and finally we attached aptamers to gold/silver nanoparticles. In addition, we developed serum stability assays to demonstrate that the RNA aptamers are not degraded by cellular nucleases thus able exert their function in live cells. We are currently testing antimicrobial effect of aptamer nanoparticle conjugates using E. coli 5922.