Determine the effect of active compounds or external factors on gene expression
Study the impact of an environment on gene activity
In many research projects, it is essential to understand how an external factor influences gene expression. An active compound, a pharmacological treatment, environmental stress, or an experimental modification can alter an organism’s transcriptional activity.
Studying these variations is based on the analysis of RNAs produced during gene transcription. By comparing expression levels across different experimental conditions, it becomes possible to identify genes whose activity is modulated by the factor under study.
This approach makes it possible to explore the molecular mechanisms involved in an organism’s biological response.
Why analyse the effect of a factor on gene expression?
Gene expression analysis makes it possible to observe an organism’s response to external stimulation. It is frequently used to:
- analyse the effect of a drug or chemical compound on the activity of certain genes
- understand an organism’s molecular response to environmental stress
- study the impact of an experimental treatment on the expression of a panel of genes
- identify biological pathways activated or inhibited under a given condition.
These analyses make it possible to link an observable biological phenomenon to specific changes in gene expression.
Principle of the analysis
Studying the effect of a molecule or an external factor generally relies on comparing several experimental conditions:
- a reference sample (control condition)
- one or more samples exposed to the factor under study.
After RNA extraction, it is converted into complementary DNA (cDNA) for analysis. Gene expression levels can then be compared across the different experimental conditions to detect exposure-induced variations.
Possible analysis methods
The choice of analytical strategy depends on the number of genes studied and the project’s scientific objectives.
Targeted analysis by RT-qPCR
Real-time quantitative PCR (RT-qPCR) makes it possible to precisely measure the expression level of a gene of interest. This method amplifies a specific genetic sequence and measures, in real time, the amount of amplified product using a fluorescent signal.
It is particularly suitable when the study focuses on:
a specific gene
a small number of targeted genes
validation of results obtained from more global analyses.
Transcriptome sequencing (RNA-Seq)
When the objective is to explore the impact of a factor on a large number of genes, it is possible to perform transcriptome sequencing (RNA-Seq).
This approach makes it possible to:
- identify the genes expressed in the sample
- measure their expression level
- detect expression changes at the transcriptome scale.
It therefore provides a global view of an organism’s genetic response.
Genetic material analysed
Depending on the study objectives, different types of genetic material may be used:
- total RNA
- total messenger RNA
- targeted mRNAs corresponding to genes of interest.
The choice of RNA type depends on the type of analysis and the desired level of precision.
An approach tailored to each scientific project
Studying the effect of active compounds or external factors on gene expression can be applied to many fields:
- molecular biology
- microbiology
- medical research
- biotechnologies
- environmental studies.
Analytical strategies can be adapted depending on the sample type, the number of genes studied, and the level of precision required.
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