Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Amuc_1100 alleviates HFD-induced hepatic lipid accumulation via gut microbiota in zebrafish: insights from the role of intestinal 14-3-3β/α-A.

Journal of animal science and biotechnology·2026
Same author

Tolerance and response of afatinib combined with tegafur in patients with advanced biliary tract cancer after failure of gemcitabine-based systemic therapy.

Discover oncology·2026
Same author

Engineering <i>Clostridium Tyrobutyricum</i> for High Butanol Production through Induction Expression of Exogenous NADPH-Dependent HBD.

ACS synthetic biology·2026
Same author

Fault Diagnosis Method for Reciprocating Compressors Based on Spatio-Temporal Feature Fusion.

Sensors (Basel, Switzerland)·2026
Same author

Akkermansia muciniphila alleviates metabolic disorders through gut microbiota-mediated tryptophan regulation.

AMB Express·2025
Same author

Programmable Anisotropic Hydrogels for Biomedicine: From Precise Design to Advanced Applications.

ACS applied materials & interfaces·2025

Related Experiment Video

Updated: May 10, 2025

High-throughput Screening for Chemical Modulators of Post-transcriptionally Regulated Genes
09:44

High-throughput Screening for Chemical Modulators of Post-transcriptionally Regulated Genes

Published on: March 3, 2015

9.4K

Development of 3D Cell-Based Fluorescent Reporter Assay for Screening of Drugs Downregulating Telomerase Reverse

You Li1, Fengli Zhang1, Zhen Qin1

  • 1William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, 151 West Woodruff Avenue, Columbus, OH 43210, USA.

Bioengineering (Basel, Switzerland)
|April 26, 2025
PubMed
Summary

A new fluorescent assay screens for cancer drugs targeting human telomerase reverse transcriptase (hTERT). This method uses EGFP reporter cells in 3D cultures, achieving over 83.3% accuracy in identifying hTERT inhibitors.

Keywords:
breast cancerdrug discoveryfluorescent reporter assayhigh-throughput screeningtelomerase reverse transcriptase

More Related Videos

A Fluorescence-based Lymphocyte Assay Suitable for High-throughput Screening of Small Molecules
08:43

A Fluorescence-based Lymphocyte Assay Suitable for High-throughput Screening of Small Molecules

Published on: March 10, 2017

10.3K
A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells
10:13

A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells

Published on: July 3, 2013

11.0K

Related Experiment Videos

Last Updated: May 10, 2025

High-throughput Screening for Chemical Modulators of Post-transcriptionally Regulated Genes
09:44

High-throughput Screening for Chemical Modulators of Post-transcriptionally Regulated Genes

Published on: March 3, 2015

9.4K
A Fluorescence-based Lymphocyte Assay Suitable for High-throughput Screening of Small Molecules
08:43

A Fluorescence-based Lymphocyte Assay Suitable for High-throughput Screening of Small Molecules

Published on: March 10, 2017

10.3K
A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells
10:13

A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells

Published on: July 3, 2013

11.0K

Area of Science:

  • Oncology
  • Molecular Biology
  • Biotechnology

Background:

  • Human telomerase reverse transcriptase (hTERT) is reactivated in over 90% of cancers, making it a key therapeutic target.
  • hTERT is essential for cancer cell proliferation and survival, but typically silenced in normal adult tissues.
  • Targeting hTERT offers a promising strategy for selective cancer treatment.

Purpose of the Study:

  • To develop and validate a novel fluorescent cell-based assay for screening compounds that inhibit hTERT expression.
  • To utilize the hTERT promoter linked to enhanced green fluorescent protein (EGFP) as a reporter for promoter activity.
  • To assess the assay's performance using known hTERT inhibitors and established anticancer drugs in a 3D culture system.

Main Methods:

  • Cloning the hTERT promoter to control EGFP expression in MCF7 breast cancer cells.
  • Utilizing a human cytomegalovirus (CMV) promoter as a control for EGFP expression.
  • Culturing EGFP reporter cells in 3D microbioreactors to mimic in vivo tumor microenvironments.
  • Evaluating assay predictability with known hTERT inhibitors (pristimerin, EGCG, n-butylidenephthalide) and other anticancer agents.

Main Results:

  • The EGFP fluorescence intensity directly correlated with hTERT promoter activity.
  • Inhibition of hTERT led to a measurable decrease in EGFP fluorescence.
  • The assay demonstrated high accuracy (over 83.3%) in identifying compounds that repress hTERT expression.
  • The 3D culture system provided in vivo-like responses for drug screening.

Conclusions:

  • The developed fluorescent assay is a feasible and accurate tool for screening potential cancer therapeutics targeting hTERT.
  • The EGFP reporter system linked to the hTERT promoter effectively monitors drug-induced changes in hTERT activity.
  • This assay, particularly within a 3D culture context, holds promise for accelerating the discovery of novel anti-cancer drugs.