Essential Role of Targeted Reporter Gene Assays in Therapeutic Assessment

Cell-based targeted reporter gene assays play a vital role in evaluating drug biological activity with high sensitivity and specificity. By placing reporter genes under specific promoters, this advanced technology allows researchers to directly determine if therapeutic agents activate or repress these pathways, revealing how compounds influence specific signaling pathways. As a result, developing reporter gene assays for multiple targets can significantly advance therapeutics development and accelerate the progression of effective therapeutics into clinical treatment.

Fig.1 A diagram of how a reporter gene is used to study a regulatory sequence. (TransControl at en.Wikipedia, CC BY-SA 3.0, https://commons.wikimedia.org/wiki/File:Reporter_gene.png)Fig.1 A diagram showing how the reporter gene works.1

Other-targeted Reporter Gene Assay Development at Creative Biolabs

Reporter gene assays offer significant advantages over traditional primary cell-based assays, including higher efficiency, consistency, and reproducibility. These assays translate candidate compound activity into measurable fluorescence, allowing real-time monitoring in living cells. Creative Biolabs leverages this technology to develop customized reporter gene assays for a variety of targets, including immune checkpoints, cytokines, growth factors, receptors, and other interest targets. Additionally, our immune cells-targeted reporter gene assays are ideal for tracking the activation of immune cells, such as T-cells and CAR-T cells, in response to immunotherapies. Of note, we provide well-validated in-house cell lines and also provide customized service to meet specific customers' needs. Creative Biolabs ensures fast, high-quality results with standardized reports delivered within 2 weeks.

Here we list several hot reporter gene assays targeting growth factors and receptors, kinase, as well as immune cells, for customers' project reference:

  • VEGF/VEGFR-targeted Reporter Gene Assay
  • ANG2-Tie2-targeted Reporter Gene Assay
  • T cell Reporter Gene Assay

Fig.2 Our assay workflow. (Creative Biolabs Original)Fig.2 Workflow of our targeted reporter gene assay.

Advantages of Targeted Reporter Gene Assay

  • High-Throughput Screening: These assays can be easily adapted to high-throughput formats, allowing the screening of thousands of compounds for desired activities.
  • Quantitative Measurement: Reporter gene assays offer a quantitative readout that can be used to measure the potency and efficacy of compounds. This is important for lead optimization and for determining the dose-response relationship.
  • Specificity and Selectivity: By linking the expression of a reporter gene to a particular cellular promoter or response element, researchers can ensure that the assay specifically measures the activation or inhibition of a target pathway, thus ensuring selectivity.
  • Live-cell Monitoring: Reporter gene assays can be designed to monitor cellular responses in real-time in living cells.
  • Versatility: Various reporter genes (e.g., luciferase, GFP, β-galactosidase) can be used depending on the detection method and sensitivity required, making the assays highly versatile.

Representative Data

Summary: TGF-β and VEGFA are key negative regulators of antitumor immunity. They are involved in immune escape and immunotherapy resistance in tumor cells. This study introduced a novel bispecific antibody that targets both TGF-β and VEGF. The biological activity of the bispecific antibody was evaluated using a series of approaches and luciferase reporter assay, respectively.
Result: The findings demonstrated that this bispecific antibody effectively inhibited both TGF-β and VEGF signaling pathways. When used in conjunction with PD-1 blockade, it showed enhanced antitumor efficacy compared to monotherapies, primarily by modulating and improving the immune microenvironment.
Fig.3 The antagonistic effect of the developed bispecific antibody on the activation of the VEGF/VEGFR pathway, VEGFA-induced proliferation and tube formation in HUVECs. (Niu, et al., 2023)
Fig.3 Evaluation of the bispecific antibody's antagonistic impact on VEGF/VEGFR pathway activation, VEGFA-driven proliferation, and tube formation in HUVECs.2, 3

If you want to customize a specific targeted reporter gene assay, please don't hesitate to reach out to us with your challenging projects.

References

  1. From Wikimedia: By TransControl at en.Wikipedia, CC BY-SA 3.0, https://commons.wikimedia.org/wiki/File:Reporter_gene.png
  2. Niu, Mengke, et al. "Synergistic efficacy of simultaneous anti-TGF-β/VEGF bispecific antibody and PD-1 blockade in cancer therapy." Journal of Hematology & Oncology 16.1 (2023): 94.
  3. under Open Access License CC BY 4.0. The original image was modified by extracting and using a part, and the title was changed to "Evaluation of the bispecific antibody's antagonistic impact on VEGF/VEGFR pathway activation, VEGFA-driven proliferation, and tube formation in HUVECs".

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