The Effects of a Novel CRISPR-Cas9 System in Human Cancer Cells

Overview

Current treatment options for lung cancer are often ineffective and nonselective, causing severe side effects and damage to healthy respiratory tissue. However, considering that cancers are caused by DNA mutations, gene therapy is now at the forefront of treatment research, and CRISPR/Cas9 may be the most accurate, fast, and cost-effective method. In this novel experiment, the effectiveness of this technology in combating tumor development was measured by using Cas9 and sgRNA molecules to target flanking ends of key exons in TERT and KRAS, intercorrelated oncogenes that promote unregulated growth and proliferation, in A549 lung cancer cells. One culture of cells was exposed to only TERT removal, another to only KRAS inactivation, and a third to targeting of both genes. Cell death rates were quantified through the trypan blue exclusion assay, while mitotic indices were calculated by observing the number of dividing cells in a constant sample size. The results supported the hypothesis, as they revealed that in comparison to the control, the groups that experienced a loss of function in both genes simultaneously consistently exhibited the highest rates of cell damage and the lowest percentages of proliferation, while those who had solely one of the genes disabled also displayed antitumoral effects, but to a lesser extent. Microscopic analysis further proved that by disrupting the expression of both oncogenes at the same time, the dual-CRISPR/Cas9 system most efficiently activated intracellular apoptotic pathways and helped the cell resist pro-mitotic environmental cues.

Competition history

• JSHS 2022 Category not listed

Resources

• National JSHS abstract book ↗

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