Posters
Presenting Author Academic/Professional Position
Medical Student
Academic Level (Author 1)
Medical Student
Discipline/Specialty (Author 1)
Immunology and Microbiology
Presentation Type
Poster
Discipline Track
Clinical Science
Abstract Type
Research/Clinical
Abstract
Many inflammatory joint diseases are associated with the expression of CD10 protein, which plays a large part in the pro-inflammatory and pain transmission signaling of the infrapatellar fat pad. This pro-inflammatory mechanism is a prime indicator in the degradation of the articular cartilage of various joints found in human musculoskeletal tissue. CD10 expression in Mesenchymal Stem Cell (MSC) is directly related to its immunomodulatory and chondroprotective effects. Thus, this project focuses on developing an aptamer-CRISPR based biosensor that will detect CD10 expression without perturbation of the sample. Aptamers are small, single-stranded nucleic acid molecules that can fold into distinctive structures, enabling them to bind with high specificity to various molecular protein targets. This allows them to detect a large array of both high and low-abundance molecules. The initial step in the project is to develop high-affinity aptamers for CD10 using a procedure referred to as SELEX (Systematic Evolution of Ligands by Exponential Enrichment). We started with an initial single-stranded RNA library that contains approximately 1014 different sequences. The RNA library was incubated with the CD10 protein in solution. The protein-RNA complex was then separated from the uncomplexed RNA using a nitrocellulose filter. We then separated the protein from the RNA, prior to subjecting the RNA to reverse transcription and PCR. The final product after the first round contains ssRNA molecules that bind to CD10 protein. This process will be repeated approximately 10 more times, allowing us to identify RNA aptamers that bind with high affinity to CD10. This is a critical step in developing the aptamer CRISPR sensor since some samples will have a low expression of CD10. I have completed 2 rounds of SELEX with promising results.
Recommended Citation
Punch, Kory; Hu, Christopher; Fortenberry, Yolanda; and Zaman, Khamequz, "Development of high affinity aptamers to CD10 Recombinant Human Neprilysin protein for aptamer-CRISPR Biosensor" (2025). Research Symposium. 91.
https://scholarworks.utrgv.edu/somrs/2025/posters/91
Included in
Development of high affinity aptamers to CD10 Recombinant Human Neprilysin protein for aptamer-CRISPR Biosensor
Many inflammatory joint diseases are associated with the expression of CD10 protein, which plays a large part in the pro-inflammatory and pain transmission signaling of the infrapatellar fat pad. This pro-inflammatory mechanism is a prime indicator in the degradation of the articular cartilage of various joints found in human musculoskeletal tissue. CD10 expression in Mesenchymal Stem Cell (MSC) is directly related to its immunomodulatory and chondroprotective effects. Thus, this project focuses on developing an aptamer-CRISPR based biosensor that will detect CD10 expression without perturbation of the sample. Aptamers are small, single-stranded nucleic acid molecules that can fold into distinctive structures, enabling them to bind with high specificity to various molecular protein targets. This allows them to detect a large array of both high and low-abundance molecules. The initial step in the project is to develop high-affinity aptamers for CD10 using a procedure referred to as SELEX (Systematic Evolution of Ligands by Exponential Enrichment). We started with an initial single-stranded RNA library that contains approximately 1014 different sequences. The RNA library was incubated with the CD10 protein in solution. The protein-RNA complex was then separated from the uncomplexed RNA using a nitrocellulose filter. We then separated the protein from the RNA, prior to subjecting the RNA to reverse transcription and PCR. The final product after the first round contains ssRNA molecules that bind to CD10 protein. This process will be repeated approximately 10 more times, allowing us to identify RNA aptamers that bind with high affinity to CD10. This is a critical step in developing the aptamer CRISPR sensor since some samples will have a low expression of CD10. I have completed 2 rounds of SELEX with promising results.
