Related Experiment Video

Updated: Jul 26, 2026

Identification of Growth Inhibition Phenotypes Induced by Expression of Bacterial Type III Effectors in Yeast
09:34

Identification of Growth Inhibition Phenotypes Induced by Expression of Bacterial Type III Effectors in Yeast

Published on: March 30, 2010

Positive selection of growth-inhibitory genes

J Z Maines1, A Sunnarborg, L M Rogers

  • 1Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis 55455, USA.

Cell Growth & Differentiation : the Molecular Biology Journal of the American Association for Cancer Research
|June 1, 1995
PubMed

Insights

Researchers identified novel growth-inhibitory genes using a unique dye retention assay. This method tracks cell division by monitoring fluorescent dye dilution, enabling the isolation of genes that limit cell proliferation.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • Identifying genes that inhibit cell proliferation is challenging due to the lack of robust screening assays.
  • Existing methods for detecting growth inhibitors are often difficult, imprecise, or incomplete.

Purpose of the Study:

  • To develop and validate a novel assay for identifying cDNAs that inhibit cell proliferation.
  • To isolate growth-inhibitory genes from epithelial cells using this new method.

Main Methods:

  • Developed a unique assay utilizing a lipophilic fluorescent dye (PKH-2) to monitor cell division.
  • Cells were labeled with the dye, transfected with a cDNA library, and sorted based on dye retention using fluorescence-activated cell sorting (FACS).
  • Growth-inhibited cells retain more dye per cell over time compared to actively dividing cells.

Main Results:

  • Successfully isolated a set of cDNAs that limit epithelial cell proliferation.
  • The dye retention assay effectively selected for pools of cDNAs with growth-inhibitory activity.
  • Demonstrated the assay's ability to identify genes that inhibit cell proliferation.

Conclusions:

  • The developed dye retention assay provides a powerful and generalizable method for identifying growth-inhibitory genes.
  • This approach can be applied across various biological contexts and cell types.
  • The study successfully identified cDNAs that inhibit epithelial cell proliferation, highlighting the assay's efficacy.

More Related Videos

Identifying Amino Acid Overproducers Using Rare-Codon-Rich Markers10:41

Identifying Amino Acid Overproducers Using Rare-Codon-Rich Markers

Published on: June 24, 2019

In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing10:44

In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing

Published on: May 5, 2023

Related Concept Videos

Feedback Inhibition00:46

Feedback Inhibition

Biochemical reactions are occurring constantly in cells, converting starting substances to different products, usually with the help of enzymes that speed the reactions. Without enzymes, it would take far too long for most reactions to occur to be useful to the cell!
Negative Regulator Molecules01:23

Negative Regulator Molecules

Positive regulators allow a cell to advance through cell cycle checkpoints. Negative regulators have an equally important role as they terminate a cell’s progression through the cell cycle—or pause it—until the cell meets specific criteria.
Antibiotic Selection00:57

Antibiotic Selection

Overview
Eukaryotic Transcription Inhibitors01:52

Eukaryotic Transcription Inhibitors

Certain biochemical processes, such as embryonic development and cell growth regulation, depend on the repression of specific genes. DNA binding proteins known as eukaryotic transcription inhibitors regulate the repression of gene expression in eukaryotes. The presence of these inhibitors at the required location and time in the cell is triggered by the presence of hormones and additional signals from other cells.
Eukaryotic transcription inhibitors usually contain two distinct domains, a DNA...
mTOR Signaling and Cancer Progression03:03

mTOR Signaling and Cancer Progression

The mammalian target of rapamycin or mTOR protein was discovered in 1994 due to its direct interaction with rapamycin. The protein gets its name from a yeast homolog called TOR. The mTOR protein complex in mammalian cells plays a major role in balancing anabolic processes such as the synthesis of proteins, lipids, and nucleotides and catabolic processes, such as autophagy in response to environmental cues, such as availability of nutrients and growth factors.
The mTOR pathway or the...
Cancer-Critical Genes II: Tumor Suppressor Genes01:05

Cancer-Critical Genes II: Tumor Suppressor Genes

Genes usually encode proteins necessary for the proper functioning of a healthy cell. Mutations can often cause changes to the gene expression pattern, thereby altering the phenotype.
When the function of certain critical genes, especially those involved in cell cycle regulation and cell growth signaling cascades, gets disrupted, it upsets the cell cycle progression. Such cells with unchecked cell cycles start proliferating uncontrollably and eventually develop into tumors.
Such genes that act...