Freedom Johnson '22
Student and Faculty Excellence (SAFE) Fund Recipient: Spring 2023
Project: "Recognition of the ClbR Gene in Escherichia coli with Hairpin Polyamide-FRET Conjugates"
Johnson received a Spring 2023 Student and Faculty Excellence (SAFE) grant to purchase the Cy3 and Cy5 chromophores needed to synthesize hairpin polyamide molecules for her project “Recognition of the ClbR Gene in Escherichia coli with Hairpin Polyamide-FRET Conjugates.”
"Hairpin polyamide-fluorescence resonance energy transfer (FRET) conjugates can signal the presence of Crohn’s disease," explained Johnson. "Hairpin polyamides designed to target the clbR gene, the key transcriptional activator essential in regulation of colibactin, a causal agent of associated inflammation in colorectal cancer, are synthesized through the covalent linkage of heterocyclic monomers, forming hairpin polyamides. These molecules are covalently linked to FRET chromophores which fluorescently signal binding to the clbR gene. If the clbR gene sequence is absent in the sample, the conjugate will not bind, fluorescence will not be observed, and Crohn’s disease is likely absent. This method offers the potential for high-throughput gene detection."
Last summer, Johnson worked toward the synthesis of hairpin polyamide FRET-conjugates and successfully constructed the first pyrrole trimer that will be used as a sequence identifier for the gene detection molecule. She also optimized the coupling step through the experimental process and found an agent for amide bond coupling to afford efficient hairpin synthesis.
"The funding from the SAFE grant enhanced my experience by learning how chemicals are chosen, found, and ordered in an organic chemistry lab," shared Johnson, who graduated from App State with her bachelor's degree in cell and molecular biology and a minor in chemistry in 2022. Currently, Johnson is pursuing her master's degree in biology. Johnson's research is advised by Dr. Rachel Bleich, professor in the Department of Biology, and Dr. Nicholas Shaw, professor in the Department of Chemistry and Fermentation Sciences.
"In the near future, experiments will be conducted with DNA samples to prove that these molecules can recognize and bind to a DNA sequence of interest, and release a recognizable signal upon binding. The release of a recognizable signal could not be done without the Cy3 and Cy5 chromophores that were purchased with the SAFE grant funds. These hairpins could be used as a new method to gene detection that is more affordable, cost-effective, and efficient," explained Johnson.
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About the SAFE Fund
Initially endowed by Hughlene and Bill Frank, the College of Arts and Sciences Student and Faculty Excellence (SAFE) Fund provides resources that can be used to support undergraduate, graduate and faculty experiences. The SAFE Fund provides funding for college priorities and opportunities that arise throughout the year. These unrestricted funds support student and faculty travel, publication support for faculty and student research opportunities. Learn more at: https://cas.appstate.edu/students/student-and-faculty-excellence-safe-fund.
About the Department of Biology
The Department of Biology is a community of teacher-scholars, with faculty representing the full breadth of biological specializations — from molecular genetics to landscape/ecosystem ecology. The department seeks to produce graduates with sound scientific knowledge, the skills to create new knowledge, and the excitement and appreciation of scientific discovery. Learn more at https://biology.appstate.edu.
About the A.R. Smith Department of Chemistry and Fermentation Sciences
The A.R. Smith Department of Chemistry and Fermentation Sciences offers a Bachelor of Arts in chemistry, a Bachelor of Science in chemistry with eight different concentrations and an interdisciplinary Bachelor of Science degree in fermentation sciences. The department’s programs prepare students to attend graduate and professional schools, as well as for employment in the pharmaceutical and fermentation industries and other business sectors. Learn more at https://dcfs.appstate.edu.
Written by Madison Breedlove
Edited by Lauren Gibbs
April 16, 2024
BOONE, N.C.
Freedom Johnson '22
Freedom Johnson '22