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Related Experiment Video

Updated: May 14, 2026

Non-Viral Engineering of Primary Human T Cells via Homology-Mediated End-Joining Targeted Integration of Large DNA Templates

Non-Viral Engineering of Primary Human T Cells via Homology-Mediated End-Joining Targeted Integration of Large DNA Templates

Published on: May 9, 2025

Multi-cistronic vector encoding optimized safety switch for adoptive therapy with T-cell receptor-modified T cells.

M M van Loenen¹, R de Boer, R S Hagedoorn

¹Department of Hematology, Leiden University Medical Center, Leiden, The Netherlands. m.m.van_loenen@lumc.nl

|February 1, 2013

Summary

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Engineered T cells for cancer therapy can be made safer with a suicide gene. Codon optimization improved the coexpression of the T-cell receptor (TCR) and safety switch (CD20) in engineered T cells, enabling their selective elimination.

Area of Science:

Immunotherapy
Cancer Biology
Molecular Engineering

Background:

T-cell receptor (TCR) gene transfer equips T cells to target cancers but carries risks like on-target toxicity and off-target reactivity.
Safety switches, such as the human CD20 gene, are crucial for controlling engineered T cells.
Previous attempts at co-expressing TCR and CD20 via multi-cistronic vectors resulted in poor coexpression.

Purpose of the Study:

To improve the coexpression of TCR and CD20 in engineered T cells for enhanced safety.
To evaluate the efficacy of codon optimization in overcoming coexpression challenges.
To demonstrate the potential of codon-optimized CD20 as a clinical safety switch.

Main Methods:

Utilized codon optimization for both TCR and CD20 genes.

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Last Updated: May 14, 2026

Non-Viral Engineering of Primary Human T Cells via Homology-Mediated End-Joining Targeted Integration of Large DNA Templates

Non-Viral Engineering of Primary Human T Cells via Homology-Mediated End-Joining Targeted Integration of Large DNA Templates

Published on: May 9, 2025

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

Constructed multi-cistronic vectors for T-cell transduction.

Assessed coexpression levels of PRAME-TCR and CD20 in engineered T cells.

Evaluated the selective elimination of engineered T cells in vitro.

Main Results:

Codon optimization significantly enhanced the coexpression of PRAME-TCR and CD20.
Engineered T cells demonstrated efficient and selective elimination in vitro.
The study successfully overcame previous coexpression limitations.

Conclusions:

Codon-optimized CD20 facilitates robust coexpression with TCRs in engineered T cells.
This approach offers a promising safety switch for TCR-based immunotherapies.
Optimized CD20 holds significant potential for clinical applications in cancer treatment.