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Updated: May 16, 2025

Real-time Analysis of Transcription Factor Binding, Transcription, Translation, and Turnover to Display Global Events During Cellular Activation

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Exploring the Perturbation of Biological Processes Caused by Gene Upregulation Using Knock-In Transfection,

Isabella R Garcia¹, Lee D Larcombe²

¹Applied Exomics Ltd (Apexomic), Stevenage, Hertfordshire, UK.

Methods in Molecular Biology (Clifton, N.J.)

|March 31, 2025

Summary

Investigating gene function is crucial for understanding biological pathways. This study combines lab experiments with computational analysis to uncover new gene roles and mechanisms of action.

Keywords:

Bioinformatics Differential expression analysis Functional enrichment analysis Gene overexpression Human embryonic kidney cells 263 (HEK293)RNA extraction Transcriptomics Transfection mRNA sequencing

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Area of Science:

Molecular Biology
Genomics
Bioinformatics

Background:

Gene overexpression in mammalian cell lines is a key method for functional genomics.
Understanding gene function is essential for dissecting complex biological pathways.

Purpose of the Study:

To describe a combined approach for gene function discovery.
To illustrate the utility of integrating wet-lab and in-silico methods.

Main Methods:

Gene overexpression in mammalian cell lines.
Wet-lab experimental techniques.
In-silico transcriptomic analysis.

Main Results:

Facilitates the discovery of previously unknown gene functions.
Enables the identification of perturbed molecular processes.
Aids in the elucidation of mechanisms of action for genes.

Conclusions:

The integration of experimental and computational approaches offers a powerful strategy for biological discovery.
This methodology enhances the investigation of gene functions and molecular mechanisms.