Exosomes, small extracellular vesicles of 30–150 nm, have moved beyond being simple mediators of intercellular communication to become next-generation therapeutic vehicles. One of the most innovative applications in recent years is their use as delivery vectors for CRISPR/Cas9 systems, offering a non-viral, safe, and highly selective approach for gene editing in diseases such as cancer and liver fibrosis.
The CRISPR/Cas9 system has revolutionized biomedicine due to its ability to perform precise DNA cuts, but its clinical application is limited by challenges in efficient delivery, cell specificity, and safety. Viral vectors, commonly used in research, carry risks of immunogenicity, mutagenesis, and complex manufacturing.
Here, exosomes emerge as a disruptive alternative:
They can be efficiently loaded with guide RNA (sgRNA) and Cas9 mRNA/protein.
They exhibit low immunogenicity, as they are derived from host cells.
Their natural tropism can be engineered to target specific cell types, such as tumor cells or hepatic stellate cells.
They can overcome physiological barriers and reach otherwise inaccessible tissues.
Recent studies have demonstrated that mesenchymal stem cell-derived exosomes loaded with CRISPR successfully edited the oncogene Kras^G12D in pancreatic cancer models, significantly reducing tumor growth (PMC8321670)
In oncology, CRISPR-functionalized exosomes target key genes involved in tumor proliferation and survival. They also modulate the tumor microenvironment that supports metastasis. This strategy enables highly specific therapies. The gene cargo can be designed according to each patient’s mutational profile.
In liver fibrosis, engineered exosomes deliver CRISPR systems that silence pro-fibrotic genes in hepatic stellate cells. This approach reverses cell activation and slows disease progression. The effect has been demonstrated in preclinical models (Frontiers in Molecular Biosciences, 2025).
These applications not only highlight therapeutic potential but also position exosomes as a central tool in precision medicine.
Despite encouraging results, several challenges remain:
Standardization of exosome loading protocols for CRISPR cargo.
Scalability of production under GMP conditions.
Long-term safety, avoiding off-target edits.
International regulation, which must adapt to these hybrid technologies at the intersection of synthetic biology and nanomedicine.
Future directions point to CRISPR-loaded exosomes being combined with immunotherapies or molecular inhibitors, creating synergistic strategies against high-mortality diseases such as pancreatic cancer or advanced fibrosis.
The convergence of exosomes and CRISPR/Cas9 represents one of the most promising frontiers in modern biomedicine. The possibility of achieving targeted, safe, and efficient gene editing through extracellular vesicles opens a horizon where personalized medicine transitions from a future projection to an actively developing reality.
In this context, advancing technologies for exosome isolation, characterization, and analysis (such as those we provide at Immunostep) will be crucial to bringing these applications closer to real-world clinical practice.