Posters
Presenting Author Academic/Professional Position
Medical Student
Academic Level (Author 1)
Medical Student
Academic Level (Author 2)
Faculty
Discipline Track
Biomedical Science
Abstract Type
Research/Clinical
Abstract
Background: Drug development research has long focused on synthesizing novel therapeutics while overlooking isolation from naturally available sources such as animal venoms. Recently, the discovery of the therapeutic effects of highly bioavailable animal venom caused the field of venom research to soar in popularity. Animal venom serves as a natural, sophisticated tool optimized over millions of years by evolution with the ability to bind to ion channels such as nAChRs, non-specific ligand-gated ion channels distributed throughout the human nervous system. However, purifying individual venom toxins from crude venom is challenging due to its availability. Therefore, synthesizing large quantities of venom toxins economically and with maximum efficiency remains a critical issue in the field of biomedical research. Herein, we clone a transcript encoding a novel peptide named P1A10 from a mixed tarantula venom gland cDNA library for recombinantly generating large quantities of venom toxin.
Methods: Four spiders of different species were grown to full size and euthanized in accordance with the Animal Use and Care Committee established by UTRGV. The venom glands were then removed and mixed to create a mixed spider venom gland from which mRNA was isolated and used to construct a cDNA library. The venom gland cDNA library was further sequenced. Full-length sequences were then translated into their amino acid sequence using Expasy translate and cross-referenced with the NCBI database to determine if the peptides were known or novel. The full-length transcripts, along with human nAChRs, were subcloned into both bacterial (E. coli) and yeast (S. cerevisae) expression vectors for recombinant protein generation as well as for protein-protein interactions. SDS-PAGE and immunoblots were used to confirm the presence of the recombinant protein, while affinity and size-exclusion chromatography were used to purify the desired recombinant proteins.
Results: The subcloning and restriction enzyme digestion procedure were successful as demonstrated by agarose gel electrophoresis. Both SDS-PAGE and immunoblot results revealed successful expression of desired spider toxin protein P1A10 and the nAChR subunits in both yeast and bacterial vectors. Yeast two-hybrid analysis confirmed a unique interaction between the b4 receptor and the novel spider venom toxin herein named as P1A10, interacting specifically within the extracellular domain of b4 and active site of P1A10. Computational modeling and docking studies were then performed to visualize the spider venom protein and a3b4 complex as it exists physiologically.
Conclusions: The literature on venom toxins as biotherapeutics is well-studied, with modern day examples including the use of captopril for hypertension and exenatide from the Gila monster for diabetes. The use of heterologous expression and recombinant protein production is a pragmatic, efficient, and inexpensive solution in the generation of large quantities of proteins for drug development, thereby enabling the production and distribution of lower-cost therapeutics. The application of recombinant venom proteins is far-reaching, with potential as monoclonal antibody treatments that can treat a variety of diseases with improved penetration capacity.
Presentation Type
Poster
Recommended Citation
Vega, Christine and Jia, Ying, "From Venom to Medicine: Harnessing Animal Toxins for Drug Discovery" (2025). Research Colloquium. 31.
https://scholarworks.utrgv.edu/colloquium/2025/posters/31
Included in
Biochemistry Commons, Computational Biology Commons, Medicine and Health Sciences Commons, Molecular Biology Commons, Other Neuroscience and Neurobiology Commons
From Venom to Medicine: Harnessing Animal Toxins for Drug Discovery
Background: Drug development research has long focused on synthesizing novel therapeutics while overlooking isolation from naturally available sources such as animal venoms. Recently, the discovery of the therapeutic effects of highly bioavailable animal venom caused the field of venom research to soar in popularity. Animal venom serves as a natural, sophisticated tool optimized over millions of years by evolution with the ability to bind to ion channels such as nAChRs, non-specific ligand-gated ion channels distributed throughout the human nervous system. However, purifying individual venom toxins from crude venom is challenging due to its availability. Therefore, synthesizing large quantities of venom toxins economically and with maximum efficiency remains a critical issue in the field of biomedical research. Herein, we clone a transcript encoding a novel peptide named P1A10 from a mixed tarantula venom gland cDNA library for recombinantly generating large quantities of venom toxin.
Methods: Four spiders of different species were grown to full size and euthanized in accordance with the Animal Use and Care Committee established by UTRGV. The venom glands were then removed and mixed to create a mixed spider venom gland from which mRNA was isolated and used to construct a cDNA library. The venom gland cDNA library was further sequenced. Full-length sequences were then translated into their amino acid sequence using Expasy translate and cross-referenced with the NCBI database to determine if the peptides were known or novel. The full-length transcripts, along with human nAChRs, were subcloned into both bacterial (E. coli) and yeast (S. cerevisae) expression vectors for recombinant protein generation as well as for protein-protein interactions. SDS-PAGE and immunoblots were used to confirm the presence of the recombinant protein, while affinity and size-exclusion chromatography were used to purify the desired recombinant proteins.
Results: The subcloning and restriction enzyme digestion procedure were successful as demonstrated by agarose gel electrophoresis. Both SDS-PAGE and immunoblot results revealed successful expression of desired spider toxin protein P1A10 and the nAChR subunits in both yeast and bacterial vectors. Yeast two-hybrid analysis confirmed a unique interaction between the b4 receptor and the novel spider venom toxin herein named as P1A10, interacting specifically within the extracellular domain of b4 and active site of P1A10. Computational modeling and docking studies were then performed to visualize the spider venom protein and a3b4 complex as it exists physiologically.
Conclusions: The literature on venom toxins as biotherapeutics is well-studied, with modern day examples including the use of captopril for hypertension and exenatide from the Gila monster for diabetes. The use of heterologous expression and recombinant protein production is a pragmatic, efficient, and inexpensive solution in the generation of large quantities of proteins for drug development, thereby enabling the production and distribution of lower-cost therapeutics. The application of recombinant venom proteins is far-reaching, with potential as monoclonal antibody treatments that can treat a variety of diseases with improved penetration capacity.
