Related Experiment Video

Updated: Jun 21, 2026

Controllable Ion Channel Expression through Inducible Transient Transfection
10:00

Controllable Ion Channel Expression through Inducible Transient Transfection

Published on: February 17, 2017

Transcriptional effects of transfection: the potential for misinterpretation of gene expression data generated from

Linda Jacobsen1, Susan Calvin, Edward Lobenhofer

  • 1Berit Biotech, Groveland, FL, USA. linda.jacobsen@beritbiotech.com

Biotechniques
|July 15, 2009
PubMed

Insights

Transfection introduces genes but can cause off-target effects. Choosing the right reagent and vector is crucial for accurate gene expression studies in cell lines like MCF7.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genomics

Background:

  • Transfection is a key technique for introducing genetic material into cells.
  • Distinguishing true biological responses from off-target effects is challenging.
  • Off-target effects can confound the interpretation of gene function studies.

Purpose of the Study:

  • To investigate and differentiate true biological effects from off-target effects in gene transfection experiments.
  • To assess the impact of transfection reagents and vectors on gene expression profiles.
  • To provide guidance on optimizing transfection protocols for accurate results.

Main Methods:

  • MCF7 breast cancer cells were transfected with reporter gene vectors and control vectors.
  • Differential gene expression analysis was performed using transcriptomics.
  • Comparison of gene expression changes induced by different transfection reagents.

Main Results:

  • Few differentially expressed transcripts were directly attributed to the reporter gene itself.
  • A significant number of transcripts were altered due to foreign DNA introduction, triggering immune-like responses.
  • Transfection reagents caused over a 10-fold difference in observed gene expression changes.

Conclusions:

  • The choice of transfection reagent and vector significantly influences observed gene expression.
  • Many observed changes may reflect cellular responses to foreign DNA rather than the introduced gene's function.
  • Careful selection and testing of transfection methods are essential for reliable gene function studies.

More Related Videos

Rapid Development of Cell State Identification Circuits with Poly-Transfection09:21

Rapid Development of Cell State Identification Circuits with Poly-Transfection

Published on: February 24, 2023

Transfecting RAW264.7 Cells with a Luciferase Reporter Gene12:58

Transfecting RAW264.7 Cells with a Luciferase Reporter Gene

Published on: June 18, 2015

Related Concept Videos

What is Gene Expression?01:42

What is Gene Expression?

Overview
Gene expression is the process in which DNA directs the synthesis of functional products, that is, proteins. Cells can regulate gene expression at various stages. It allows organisms to generate different cell types and enables cells to adapt to internal and external factors.
Genetic Information Flows from DNA to RNA to Protein
A gene is a stretch of DNA that serves as the blueprint for functional RNAs and proteins. Since DNA is made up of nucleotides and proteins consist of amino...
DNA-only Transposons02:57

DNA-only Transposons

DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
The donor site from where the transposon is excised is either degraded or...
Reporter Genes02:11

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.
Commonly used reporter...