CRISPR-Based Single-Nucleotide Editing of PIK3CA c.3140A>G (p.His1047Arg) in MCF7 Breast Cancer Cells Enhances Proliferative Potential.
Year: 2025
Type: article
Abstract: Objective: c.3140A>G mutation using CRISPR-Cas9 with precise single-nucleotide editing, and evaluated its impact on cellular characteristics. Materials and Methods: H1047R mutant MCF7 cells were generated using CRISPR-Cas9-mediated genome editing followed by hierarchical single-cell isolation. Editing efficiency was validated through allele-specific polymerase chain reaction (PCR) and multiple rounds of Sanger sequencing. Cell cycle distribution and proliferation were analyzed using flow cytometry and cell count assays, respectively. Gene expression changes were assessed by quantitative real-time PCR to evaluate the mutation's impact on cell cycle-related genes. Results: and MYC expression increasing by 1.62-fold (P<0.001) and 1.23-fold (P<0.001), respectively, relative to controls. Conclusion: The genetically edited cell lines represent robust and well-defined experimental models that enable direct assessment of the functional consequences of oncogenic driver mutations on cellular behavior and signaling pathways. Our findings demonstrate that targeted genetic alterations induce measurable changes in proliferation, cell-cycle regulation, and gene expression, thereby providing mechanistic insight into tumorigenesis and the specific contribution of driver mutations to cancer-related cellular phenotypes.
Source: PubMed
Institution: Tarbiat Modares University
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W7129088470
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