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Related Experiment Video

Updated: Sep 20, 2025

Author Spotlight: Efficient CRISPR/Cas9 Genome Editing in Bone Marrow-Derived Macrophages for Precise Gene Disruption

Author Spotlight: Efficient CRISPR/Cas9 Genome Editing in Bone Marrow-Derived Macrophages for Precise Gene Disruption

Published on: August 4, 2023

Engineering Human Circulating Monocytes/Macrophages by Systemic Deliverable Gene Editing.

So Yoon Lee¹, Javier Fierro¹, Jake Dipasquale¹

¹Department of Molecular and Translational Medicine of Paul L. Foster School of Medicine, Texas Tech University Health Sciences Center at El Paso, El Paso, TX, United States.

Frontiers in Immunology

|June 6, 2022

Summary

This summary is machine-generated.

We developed novel nanoparticles to deliver plasmid DNA for macrophage gene editing. This breakthrough enables effective gene delivery into macrophages, offering a new therapeutic approach for various diseases.

Keywords:

IL-4 human circulating monocytes-derived macrophages immune regulation plasmid DNA systemic deliverable gene-editing

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Last Updated: Sep 20, 2025

Author Spotlight: Efficient CRISPR/Cas9 Genome Editing in Bone Marrow-Derived Macrophages for Precise Gene Disruption

Author Spotlight: Efficient CRISPR/Cas9 Genome Editing in Bone Marrow-Derived Macrophages for Precise Gene Disruption

Published on: August 4, 2023

Preparation and Gene Modification of Nonhuman Primate Hematopoietic Stem and Progenitor Cells

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Published on: February 15, 2019

CRISPR/Cas9 Gene Editing of Hematopoietic Stem and Progenitor Cells for Gene Therapy Applications

CRISPR/Cas9 Gene Editing of Hematopoietic Stem and Progenitor Cells for Gene Therapy Applications

Published on: August 9, 2022

Area of Science:

Biomedical Engineering
Cell Biology
Gene Therapy

Background:

Macrophage genetic engineering is crucial for treating diseases like cancer and infections.
Plasmid DNA (pDNA) delivery into terminally differentiated macrophages is challenging due to nuclear membrane barriers.

Purpose of the Study:

To develop an effective method for delivering plasmid DNA into macrophages for genetic engineering.
To engineer macrophages using a novel nanoparticle (NP) system for enhanced gene delivery.

Main Methods:

Developed core-shell nanoparticles (NPs) using a novel cationic lipid to encapsulate IL-4 pDNA (IL-4pDNA-NPs).
Transfected human blood monocyte-derived macrophages (MDM) with IL-4pDNA-NPs.
Assessed NP internalization, nuclear delivery, exogenous IL-4 expression, and M2 macrophage polarization in vitro and in vivo.

Main Results:

IL-4pDNA-NPs were efficiently internalized by MDM and delivered to the nucleus within 2 hours.
Exogenous IL-4 expression was detected within 1-2 days and persisted for up to 30 days.
Functional IL-4 expression induced M2 macrophage polarization in vitro and in a mouse model of inflammation.

Conclusions:

The developed NPs protect pDNA from degradation and facilitate nuclear transport, enhancing gene delivery in macrophages.
This nanoparticle-mediated gene delivery system offers a promising strategy for macrophage-based gene therapy.
The approach provides a new solution for gene editing in monocytes and macrophages for therapeutic applications.