Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

MicroRNAs01:22

MicroRNAs

3.8K
MicroRNA (miRNA) are short, regulatory RNA transcribed from introns (non-coding regions of a gene) or intergenic regions (stretches of DNA present between genes). Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself, forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA...
3.8K
MicroRNAs01:22

MicroRNAs

23.9K
MicroRNA (miRNA) are short, regulatory RNA transcribed from introns—non-coding regions of a gene—or intergenic regions—stretches of DNA present between genes. Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After...
23.9K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Ten-Year Outcomes after CAR T-Cell Therapy for B-Cell Lymphomas.

The New England journal of medicine·2026
Same author

Single-day nonactivated IL-18-armed CAR T cells establish a durable, stemlike state with enhanced persistence.

Blood·2026
Same author

HLA-E single-chain trimer shields gene-edited allogeneic CAR T cells from NK cell attack.

Blood advances·2026
Same author

Clinicians' cancer risk assessment among patients with pulmonary nodules: a qualitative study.

Annals of the American Thoracic Society·2026
Same author

Predictive biomarkers of response to chimeric antigen receptor (CAR) T-cell therapy for pan-haematologic cancer.

Nature biomedical engineering·2026
Same author

Advancing Cell Therapies for Solid Tumors: A Pathway to Overcome Biological, Operational, and Regulatory Hurdles.

Transplantation and cellular therapy·2026

Related Experiment Video

Updated: Jan 18, 2026

Non-Viral Engineering of Primary Human T Cells via Homology-Mediated End-Joining Targeted Integration of Large DNA Templates
06:10

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

Published on: May 9, 2025

906

Multiplex engineering using microRNA-mediated gene silencing in CAR T cells.

Giulia Golinelli1,2, John Scholler1, Audrey Roussel-Gervais3

  • 1Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, United States.

Frontiers in Immunology
|September 8, 2025
PubMed
Summary
This summary is machine-generated.

Targeted microRNAs (miRNAs) offer a safer alternative to gene editing for CAR T-cell therapies, enhancing antitumor activity, persistence, and immune evasion without DNA risks.

Keywords:
CAR TCRISPR/Cas9gene silencingmiRNAsmultiplexingsolid tumors

More Related Videos

A Nonviral Approach to Generate Transient Chimeric Antigen Receptor T Cells Using mRNA for Cancer Immunotherapy
09:56

A Nonviral Approach to Generate Transient Chimeric Antigen Receptor T Cells Using mRNA for Cancer Immunotherapy

Published on: February 21, 2025

1.3K
Author Spotlight: Enhancing CAR-T Cell Function in Syngeneic Tumor Models
06:22

Author Spotlight: Enhancing CAR-T Cell Function in Syngeneic Tumor Models

Published on: February 2, 2024

4.8K

Related Experiment Videos

Last Updated: Jan 18, 2026

Non-Viral Engineering of Primary Human T Cells via Homology-Mediated End-Joining Targeted Integration of Large DNA Templates
06:10

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

Published on: May 9, 2025

906
A Nonviral Approach to Generate Transient Chimeric Antigen Receptor T Cells Using mRNA for Cancer Immunotherapy
09:56

A Nonviral Approach to Generate Transient Chimeric Antigen Receptor T Cells Using mRNA for Cancer Immunotherapy

Published on: February 21, 2025

1.3K
Author Spotlight: Enhancing CAR-T Cell Function in Syngeneic Tumor Models
06:22

Author Spotlight: Enhancing CAR-T Cell Function in Syngeneic Tumor Models

Published on: February 2, 2024

4.8K

Area of Science:

  • Immunology
  • Cell Therapy
  • Molecular Biology

Background:

  • Multiplex gene-edited CAR T-cell therapies face challenges, including oncogenic risks from DNA double-strand breaks.
  • Targeted microRNAs (miRNAs) present a safer, tunable alternative for gene silencing, avoiding DNA editing.
  • This study explores miRNA-based multiplex gene silencing in CAR T cells.

Purpose of the Study:

  • To demonstrate multiplex gene silencing using an optimized miRNA approach in CAR T cells.
  • To compare the efficacy and safety of miRNA-mediated gene silencing against CRISPR/Cas9 gene editing.
  • To evaluate the antitumor activity and immune evasion capabilities of miRNA-silenced CAR T cells.

Main Methods:

  • Engineered miRNA-expressing cassettes into M5CAR lentiviral vectors for multiplex gene silencing (TCR, MHC-I).
  • Compared miRNA-silenced (S) M5CAR T cells with CRISPR/Cas9 knockout (KO) M5CAR T cells.
  • Assessed antitumor activity in vitro and in vivo in pancreatic ductal adenocarcinoma models.

Main Results:

  • Silenced M5CAR T cells exhibited comparable or superior antitumor functionality to KO cells.
  • In vivo studies showed enhanced tumor control, persistence, and metastasis prevention with S M5CAR T cells.
  • In vitro assays revealed increased resistance to alloreactive NK cells and PBMCs.

Conclusions:

  • Titratable multiplex gene silencing via miRNAs provides a viable alternative to gene editing for CAR T cells.
  • This miRNA strategy offers potential advantages in potency, persistence, metastasis prevention, and immune evasion.
  • It may overcome tumor-induced immunosuppression while mitigating risks associated with DNA double-strand breaks.