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Related Experiment Video

Updated: Feb 7, 2026

In Vivo Direct Reprogramming of Resident Glial Cells into Interneurons by Intracerebral Injection of Viral Vectors

In Vivo Direct Reprogramming of Resident Glial Cells into Interneurons by Intracerebral Injection of Viral Vectors

Published on: June 17, 2019

Reprogramming human T cell function and specificity with non-viral genome targeting.

Theodore L Roth^1,2,3,4,5, Cristina Puig-Saus⁶, Ruby Yu^3,4,5

¹Medical Scientist Training Program, University of California, San Francisco, San Francisco, CA, USA.

|July 12, 2018

Summary

This summary is machine-generated.

This study introduces a non-viral CRISPR-Cas9 genome editing system for efficient T cell reprogramming. This method enables precise large DNA insertions for therapeutic applications, including autoimmune disease correction and cancer immunotherapy.

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Genome-wide Analysis using ChIP to Identify Isoform-specific Gene Targets

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In Vivo Direct Reprogramming of Resident Glial Cells into Interneurons by Intracerebral Injection of Viral Vectors

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Published on: June 17, 2019

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

Genome-wide Analysis using ChIP to Identify Isoform-specific Gene Targets

Genome-wide Analysis using ChIP to Identify Isoform-specific Gene Targets

Published on: July 7, 2010

Area of Science:

Immunology
Molecular Biology
Gene Therapy

Background:

Traditional T cell reprogramming relies on viral vectors, which are costly and time-consuming.
Viral vectors lack precise genomic integration, posing challenges for therapeutic applications.
Genome editing offers targeted gene insertion but has faced limitations with large DNA sequences.

Purpose of the Study:

To develop a non-viral CRISPR-Cas9 system for efficient, site-specific insertion of large DNA sequences into primary human T cells.
To demonstrate the therapeutic potential of this system in correcting genetic defects and engineering T cells for cancer immunotherapy.

Main Methods:

Developed a CRISPR-Cas9 genome-targeting system for non-viral, homology-directed repair-mediated insertion of large DNA sequences (>1 kb).
Applied the system to primary human T cells, assessing cell viability, function, and precise genomic integration.
Utilized the system to correct an IL2RA mutation in autoimmune disease models and engineer T cells with a cancer-targeting T cell receptor (TCR).

Main Results:

Achieved rapid and efficient insertion of large DNA sequences into primary human T cells without viral vectors, preserving cell viability and function.
Successfully corrected a pathogenic IL2RA mutation, restoring signaling function in T cells from patients with monogenic autoimmune disease.
Engineered T cells with a novel TCR that specifically recognized tumor antigens, demonstrating effective anti-tumor responses in vitro and in vivo.

Conclusions:

The non-viral CRISPR-Cas9 genome targeting system enables rapid and flexible genetic engineering of primary human immune cells.
This technology holds significant preclinical promise for developing novel cell-based therapies for autoimmune diseases and cancer.
Non-viral genome targeting offers a more efficient and potentially cost-effective alternative to viral vectors for T cell-based therapeutics.