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Reporter Genes

Reporter Genes

Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.

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Generation of a pHSPA6 gene-based multifunctional live cell sensor.

Zicong Xie¹, Ruize Sun¹, Chunyun Qi¹

¹Key Laboratory of Zoonosis Research, Ministry of Education, Jilin Provincial Key Laboratory of Animal Embryo Engineering, College of Animal Sciences, Jilin University, 130062 Changchun, Jilin Province, People's Republic of China.

Biochimica Et Biophysica Acta. Molecular Cell Research

|December 6, 2020

Summary

Researchers developed a novel HSPA6-based EGFP fluorescent reporter cell line using CRISPR/Cas9. This intelligent whole-cell biosensor rapidly converts cellular stress signals into detectable EGFP fluorescence.

Keywords:

CRISPR/Cas9 HSPA6 gene Knock-in Live cell sensor Stress response

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Area of Science:

Cellular biology
Biosensor technology
Gene editing

Background:

Intact live-cell biosensors offer rapid and sensitive detection of stimuli.
The HSPA6 gene's expression patterns are key to developing advanced whole-cell sensors.
Existing biosensors can be improved for greater functionality and intelligence.

Purpose of the Study:

To design and develop a novel pHSPA6-based EGFP fluorescent reporter cell line.
To create a precise and marker-free reporter system for HSPA6 expression.
To establish an intelligent live-cell sensor for stress response detection.

Main Methods:

CRISPR/Cas9-mediated knock-in strategy for precise EGFP integration.
Development of a reporter cell line with controlled gene copy number.
Stimulation experiments using HSPA6-specific stressors.

Main Results:

The pHSPA6-EGFP reporter cell line was successfully constructed with precise integration and copy number.
Reporter cells demonstrated rapid and effective conversion of stress signals into EGFP fluorescence.
Cell proliferation was normal, with sensitive and rapid expression of integrated EGFP and pHSPA6 genes upon stimulation.

Conclusions:

A new strategy for constructing HSPA6 expression/interaction cell models and intelligent live cell sensors was established.
The developed reporter cell line offers a sensitive and rapid method for detecting HSPA6-related cellular responses.
Potential applications include cellular signaling studies, biomaterials, food safety, environmental monitoring, and drug screening.