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High School Student Local to the Baltimore Area

HS Poster

Poster 114 - Molecular Cloning Techniques and Caspase-3 in Apoptosis

Saturday, October 26, 2024
10:00 AM - 12:00 PM EST
Location: Exhibit Hall E, F & G

    Presenting Author(s)

  • Sophia Yang

    Student
    Urbana High School
    Frederick, Maryland, United States

  • CL

    Claire Li

    Student
    Shadow Creek High School, United States

    • Last Author(s)

  • EC

    Elizabeth Chung

    Executive Director
    Asian American Center of Frederick, United States

    • Primary Investigator(s)

  • ZT

    Zhen Tao

    CEO
    GenuINCare, United States


Introduction: Molecular cloning techniques are crucial across numerous fields of biological and biomedical sciences. They enable the isolation, manipulation, and study of specific genes, facilitating understanding of gene function; specifically, of regulatory mechanisms and interactions between genes and proteins. Apoptosis, often referred to as programmed cell death, is a highly regulated process of cell suicide that occurs in multicellular organisms. Understanding the mechanisms of apoptosis has significant implications in medicine, particularly in cancer treatment, where inducing apoptosis in cancer cells is a key strategy for therapeutic intervention. Caspase-3 is a crucial enzyme involved in the execution phase of apoptosis and serves as a key executioner in the apoptotic cascade. Our goal was to clone Caspase-3 cDNA into a plasmid so this plasmid may induce apoptosis in cancer cells.

Materials and

Methods: PCR (Polymerase Chain Reaction) was used; PCR is a method for amplifying specific DNA sequences. Enzyme digestion with the enzymes EcoRI and XhoI was then used to precisely cut DNA at specific sequences. DNA gel electrophoresis was utilized to separate and analyze these DNA fragments based on their size. Ligation techniques were applied afterwards to join the DNA fragments that had been digested. Transformation, the process of introducing recombinant DNA into competent cells, was additionally used. Plasmid extraction with a plasmid mini prep kit was performed to isolate plasmid from competent cells. These techniques, all taught and supervised by an experienced professional, are foundational in molecular biology research, and enabled us to conduct experiments with Caspase-3 genes.

Results, Conclusions, and Discussions: Research is ongoing. Currently, we have obtained positive clones that contain Caspase-3 cDNA plasmid, as confirmed by enzyme digestion. Our future goal is to transfect the Caspase-3 gene into cancer cells to induce apoptosis. From this project, we have mastered molecular cloning and manipulation techniques, which will help us to elucidate the gene functions in regulating apoptosis in cancer cells. These techniques allow us to understand disease pathways. Our work contributes to the biomedical field of cancer research.

Acknowledgements (Optional): We would like to thank the Asian American Center of Frederick (AACF) for a grant to fund our research, and GenuINCare for allowing us to utilize their facilities.

References (Optional):