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Related Experiment Video

Updated: Apr 25, 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

Engineered mRNA backbones for gene expression in human T cells.

Gilad Gibor¹, Neve Tzvi¹, Amilia Meir²

¹Ella Lemelbaum Institute for Immuno-oncology, Sheba Medical Center, Tel Hashomer, Ramat Gan, Israel.

Molecular Therapy. Nucleic Acids

|April 24, 2026

Summary

This summary is machine-generated.

Researchers developed new messenger RNA (mRNA) systems for cancer immunotherapy by replacing standard sequences with T cell-specific ones. This optimization enhances CAR-T cell therapy effectiveness and reduces side effects.

Keywords:

CAR-T MT: Oligonucleotides: Therapies and Applications T cell biology T cell engineering TIGIT UTR immunotherapy mRNA

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Last Updated: Apr 25, 2026

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

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Published on: May 9, 2025

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Streamlined Single Cell TCR Isolation and Generation of Retroviral Vectors for In Vitro and In Vivo Expression of Human TCRs

Streamlined Single Cell TCR Isolation and Generation of Retroviral Vectors for In Vitro and In Vivo Expression of Human TCRs

Published on: September 10, 2017

Area of Science:

Immunology
Molecular Biology
Biotechnology

Background:

Current messenger RNA (mRNA) immuno-oncology therapies lack T cell specificity.
Tailored mRNA expression systems are needed for improved T cell targeting.

Purpose of the Study:

To develop novel mRNA constructs with T cell-specific 5' untranslated regions (UTRs).
To evaluate the impact of these UTRs on protein expression and CAR-T cell functionality.

Main Methods:

Replaced the standard α-globin (HBA1) 5' UTR with sequences from highly expressed T cell genes.
Assessed reporter gene and CD19-CAR construct expression in primary human T cells and HEK cells.
Evaluated protein expression, T cell function, and tonic signaling.

Main Results:

UTRs from interferon gamma (IFN-γ) enhanced protein expression, while TNF UTRs diminished it.
Expression differences were T cell-specific and did not correlate with predicted RNA stability.
The TIGIT 5' UTR optimized CD19-CAR expression, maximizing reactivity and minimizing tonic signaling.

Conclusions:

T cell-specific 5' UTRs are crucial for optimizing CAR-T cell functionality.
Fine-tuning mRNA expression via UTRs can reduce tonic signaling and off-target effects.
This approach holds promise for advancing mRNA-based CAR-T cell therapies.