Development of a novel reporter gene vector for cell based angiogenic studies
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Published on: July 30, 2018
Srinivasan Kokatam1, Deepshikha Pandey, Ramkrishna Gupta
1Premas Biotech Pvt. Ltd., Plot No. 77, Sector 4, IMT Manesar, Gurgaon, Haryana, India.
Abstract:
Angiogenesis is a promising area of research that targets key therapeutic areas like cancer; wound healing, inflammatory diseases, etc. There is an increasing demand for screening of potential angiogenic and anti-angiogenic agents using sensitive, robust cell-based assays. We have developed a reporter vector containing cis-acting elements that respond to growth factors/angiogenic ligands for use in a cell-based luciferase reporter assay. We performed transient transfection of our reporter gene vector in MCF-7 cells to establish its application for screening of potential pro/anti-angiogenic agents. Reporter gene transactivation studies with different concentrations of fetal bovine serum clearly indicated that the vector is functionally responsive to the angiogenic signals mediated by serum growth factors. We also used endostatin to inhibit transactivation and prove responsiveness to the anti-angiogenic agent. This vector is a promising tool for studying angiogenesis using cell-based reporter gene assays.
Insights
Researchers developed a novel reporter vector for cell-based assays to screen for agents that promote or inhibit angiogenesis. This tool aids in studying angiogenesis for therapeutic applications like cancer treatment.
Area of Science:
- Biotechnology
- Molecular Biology
- Cell Biology
Background:
- Angiogenesis, the formation of new blood vessels, is crucial in diseases like cancer and wound healing.
- There's a growing need for effective cell-based assays to screen for angiogenic and anti-angiogenic compounds.
- Current screening methods require robust and sensitive tools to identify therapeutic agents targeting angiogenesis.
Purpose of the Study:
- To develop and validate a novel reporter vector for a cell-based luciferase assay.
- To assess the vector's responsiveness to angiogenic and anti-angiogenic stimuli.
- To establish a tool for screening potential pro-angiogenic and anti-angiogenic agents.
Main Methods:
- A reporter vector with cis-acting elements responsive to angiogenic ligands was constructed.
- MCF-7 cells were transiently transfected with the reporter gene vector.
- Reporter gene transactivation was measured using a luciferase assay under varying conditions.
Main Results:
- The reporter vector demonstrated functional responsiveness to angiogenic signals from fetal bovine serum.
- Endostatin, an anti-angiogenic agent, successfully inhibited reporter gene transactivation, confirming vector sensitivity.
- The assay effectively screened for both pro-angiogenic and anti-angiogenic activities.
Conclusions:
- The developed reporter vector is a valuable tool for studying angiogenesis.
- This cell-based luciferase reporter assay provides a sensitive and robust method for screening therapeutic agents.
- The vector facilitates research in angiogenesis-related diseases, including cancer and inflammatory conditions.
