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Related Experiment Video

Updated: Feb 22, 2026

An Efficient In Vitro Transposition Method by a Transcriptionally Regulated Sleeping Beauty System Packaged into an Integration Defective Lentiviral Vector

An Efficient In Vitro Transposition Method by a Transcriptionally Regulated Sleeping Beauty System Packaged into an Integration Defective Lentiviral Vector

Published on: January 12, 2018

Transient and stable vector transfection: Pitfalls, off-target effects, artifacts.

Aleksei A Stepanenko¹, Henry H Heng²

¹Laboratory of Biosynthesis of Nucleic Acids, Department of Functional Genomics, Institute of Molecular Biology and Genetics, Zabolotnogo Str. 150, Kyiv 03680, Ukraine.

Mutation Research. Reviews in Mutation Research

|September 21, 2017

Summary

Vector transfection controls may not be reliable. Experimental manipulations, including empty vectors, can cause unintended genomic and phenotypic changes, impacting gene function studies.

Keywords:

Aneuploidy Chromosomal instability Geneticin/G418 Puromycin Tumor heterogeneity Zeocin

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Production of Replication-Defective Retrovirus by Transient Transfection of 293T cells

Production of Replication-Defective Retrovirus by Transient Transfection of 293T cells

Published on: December 4, 2007

Area of Science:

Molecular Biology
Genetics
Cell Biology

Background:

Vector transfection is crucial for studying gene and protein functions.
The assumption that empty vectors serve as adequate controls is widely accepted.
Unexpected results in transfection experiments challenge this assumption.

Purpose of the Study:

To investigate the validity of empty vector-transfected cells as controls in transfection experiments.
To highlight potential confounding factors introduced by transfection procedures.
To explain unexpected findings using the genome theory of somatic evolution.

Main Methods:

Review of studies reporting genomic, transcriptomic, and phenotypic changes after empty vector transfection.
Application of the genome theory of somatic evolution to explain stress-mediated dynamics.
Conceptualization of experimental manipulations as system stressors.

Main Results:

Empty vector transfection can induce significant genomic, transcriptomic, and phenotypic alterations.
Transfection reagents, antibiotic selection, and vector components can elicit cellular responses.
These changes can affect biochemical processes independently of the targeted gene.

Conclusions:

The use of empty vector-transfected cells as universal controls is not universally valid.
Experimental manipulations can act as stressors, leading to genome-level alterations (e.g., chromosomal instability).
Gene-focused methodologies may have reduced specificity due to these system-wide effects.