CRISPR/Cas9 Nanodelivery: Advancing Precision Genome

Abstract

Introduction

The CRISPR/Cas9 system has emerged as a revolutionary genome editing tool, allowing precise, efficient, and cost-effective modifications in DNA sequences. Its applications span medicine, agriculture, and biotechnology, including disease therapy, crop improvement, and livestock genetic enhancement. However, delivering CRISPR/Cas9 components—Cas9 protein and guide RNA—into target cells efficiently and safely remains a significant challenge. Nanodelivery systems have been developed to address these issues, offering protection, targeted delivery, and controlled release of CRISPR components [1]. Nanotechnology-based delivery enhances genome editing precision while minimizing off-target effects and immune responses [2].

Discussion

Effective CRISPR/Cas9 delivery is critical for successful genome editing. Traditional methods such as viral vectors, electroporation, or microinjection have limitations, including immunogenicity, insertional mutagenesis, low efficiency in certain cell types, and high technical complexity. Nanodelivery offers a versatile, non-viral alternative capable of overcoming these challenges [3].

Types of nanocarriers used for CRISPR/Cas9 delivery include lipid nanoparticles (LNPs), polymeric nanoparticles, gold nanoparticles, and dendrimers. Lipid nanoparticles, for instance, encapsulate Cas9 mRNA and guide RNA, facilitating efficient cellular uptake through endocytosis and promoting release into the cytoplasm. Gold nanoparticles and polymer-based carriers can be functionalized with targeting ligands, improving tissue-specific delivery and reducing off-target editing. Dendrimers, with their branched structure, provide high loading capacity and protect CRISPR components from degradation [4].

Targeted delivery is a major advantage of nanocarriers. Functionalization with antibodies, peptides, or aptamers enables selective binding to specific cell types, such as cancer cells or stem cells, reducing unintended genome edits in non-target tissues. Stimuli-responsive nanocarriers, sensitive to pH, temperature, or enzymatic activity, allow controlled release of CRISPR components, further enhancing editing precision.

Applications of CRISPR/Cas9 nanodelivery are vast. In medicine, it has been used to correct genetic mutations in diseases such as sickle cell anemia, Duchenne muscular dystrophy, and certain cancers. In agriculture and livestock, nanodelivery facilitates precise genome edits to improve disease resistance, productivity, and stress tolerance without introducing foreign DNA, addressing regulatory and public acceptance concerns [5].

Despite its potential, challenges remain. Nanocarrier design must balance biocompatibility, stability, and cellular uptake while minimizing cytotoxicity and immune responses. Manufacturing reproducibility, scalability, and cost-effectiveness are critical for clinical and commercial applications. Furthermore, off-target effects, although reduced by nanodelivery, must be rigorously assessed through molecular analyses and safety studies.

Conclusion

CRISPR/Cas9 nanodelivery represents a transformative approach to precise genome editing, combining the specificity of CRISPR technology with the versatility and targeting capabilities of nanocarriers. By improving cellular uptake, protecting CRISPR components, and enabling controlled and tissue-specific delivery, nanodelivery systems enhance the efficiency, safety, and applicability of genome editing across medicine, agriculture, and biotechnology. While challenges related to biocompatibility, scalability, and off-target effects persist, ongoing advances in nanotechnology and molecular biology are rapidly expanding its potential. CRISPR/Cas9 nanodelivery promises to revolutionize therapeutic interventions, crop improvement, and animal biotechnology, bringing genome editing closer to safe, precise, and practical applications.

References

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