Journal of Nanomedicine & Nanotechnology

Nanoparticle-Enhanced Gene Delivery for CRISPR-Based Therapeutics

Kjell Osterberg^*# Department of Nanomedicine, Korea Advanced Institute of Science and Technology, Korea, Democratic People's Republic of
^#Equally contribution ^*Correspondence: Kjell Osterberg, Department of Nanomedicine, Korea Advanced Institute of Science and Technology, Korea, Democratic People's Republic of, Email:

Received: 02-Nov-2024, Manuscript No. jnmnt-24-28255; Editor assigned: 05-Nov-2024, Pre QC No. jnmnt-24-28255 (PQ); Reviewed: 20-Nov-2024, QC No. jnmnt-24-28255; Revised: 25-Nov-2024, Manuscript No. jnmnt-24-28255 (R); Published: 30-Nov-2024, DOI: 10.35248/2157-7439.24.15.759

INTRODUCTION

The advent of the CRISPR-Cas9 gene-editing technology has provided unprecedented opportunities for targeted genetic modifications. CRISPR-based therapeutics hold the potential to treat a variety of diseases, including inherited genetic disorders, cancers, and viral infections, by directly modifying the genome of patient cells. However, the success of CRISPR-based therapies is contingent upon the efficient and precise delivery of the CRISPR components (Cas9 proteins, guide RNAs (gRNAs), and donor DNA) into the target cells. Conventional delivery methods, such as viral vectors (adenoviruses, lentiviruses, and retroviruses), have limitations, including the risk of immune responses, potential mutagenesis, and the complexity of large-scale manufacturing. These challenges have led to the exploration of non-viral delivery systems, with nanoparticles emerging as a promising alternative. Nanoparticles can be engineered to deliver CRISPR components in a controlled and targeted manner, improving the overall efficiency and safety of gene editing. This review examines the types of nanoparticles used in CRISPR-based therapeutics, their mechanisms of action, and the advancements in nanoparticle-enhanced gene delivery for therapeutic applications [1].

TYPES OF NANOPARTICLES FOR CRISPR DELIVERY

Nanoparticles used for CRISPR delivery are typically composed of materials that offer biocompatibility, stability, and the ability to protect the CRISPR components from degradation during delivery. The main types of nanoparticles used for CRISPR-based gene delivery include

Liposomes
Liposomes are lipid-based nanoparticles that can encapsulate both hydrophobic and hydrophilic CRISPR components, such as Cas9 proteins, gRNAs, and donor DNA. Their structure, consisting of a lipid bilayer, allows for the effective encapsulation of these components and provides protection from enzymatic degradation in vivo. Liposomes are widely used for drug delivery due to their biocompatibility and ease of functionalization with targeting ligands. In the context of CRISPR delivery, liposomes can be modified with PEGylation (polyethylene glycol) to improve their stability and circulation time in the bloodstream, as well as with cell-specific ligands to enhance targeting to specific cell types [2].

Polymeric Nanoparticles
Polymeric nanoparticles, including polymeric micelles, nanocapsules, and nanogels, offer a versatile platform for CRISPR delivery. These nanoparticles can be synthesized from biodegradable and biocompatible polymers, such as polyethyleneimine (PEI), polylactic-co-glycolic acid (PLGA), and chitosan. Polymeric nanoparticles can be easily modified to carry the CRISPR components, improve stability, and enhance delivery efficiency. Additionally, their surface can be functionalized with specific targeting molecules (e.g., antibodies, peptides) to enable cell-specific delivery. Polymeric nanoparticles also allow for controlled release of the CRISPR components, which is crucial for minimizing toxicity and improving therapeutic efficacy [3].

Inorganic Nanoparticles
Inorganic nanoparticles, including gold nanoparticles (AuNPs), silica nanoparticles, and carbon-based nanoparticles (such as graphene oxide (GO) and carbon nanotubes (CNTs)), have been increasingly explored for gene delivery applications. Gold nanoparticles, for example, can efficiently bind to gRNAs and Cas9 proteins through thiol-gold interactions, forming stable nanoparticle-CRISPR complexes that protect the gene-editing machinery from degradation. Graphene oxide is another promising material due to its high surface area, easy functionalization, and ability to encapsulate CRISPR components. The large surface area of these inorganic nanoparticles allows for the loading of a higher amount of CRISPR cargo, making them highly efficient for gene delivery [4].

Nanodiamonds
Nanodiamonds (NDs), which are diamond-like carbon nanoparticles, are gaining attention for CRISPR delivery due to their biocompatibility, low toxicity, and ability to bind to various biomolecules. Nanodiamonds have functional groups on their surface that can be modified to interact with CRISPR components, allowing for effective gene delivery. NDs can also be used for multifunctional delivery systems by integrating imaging and therapeutic agents on the same particle, providing a powerful platform for both CRISPR gene editing and tracking in vivo [5].

MECHANISMS OF NANOPARTICLE-MEDIATED CRISPR DELIVERY

The efficient delivery of CRISPR components requires overcoming several biological barriers, including cellular uptake, endosomal escape, and intracellular release of the CRISPR components. Nanoparticles enhance gene delivery through various mechanisms

Cellular Uptake
Nanoparticles can be internalized by cells through endocytosis or other mechanisms, such as clathrin-mediated or caveolae-mediated pathways. The small size of nanoparticles enables efficient uptake by cells, even by hard-to-transfect cell types. Surface modifications, such as the addition of targeting ligands (e.g., antibodies or peptides), can enhance the specificity of nanoparticle uptake by certain cell types, ensuring that CRISPR components are delivered to the desired cells [6].

Endosomal Escape
Once internalized by cells, nanoparticles must escape from the endosomal compartments to release the CRISPR components into the cytoplasm. This is a significant challenge for many non-viral gene delivery systems. To overcome this barrier, nanoparticles are often designed with endosomal escape mechanisms, such as the incorporation of cationic materials (e.g., PEI) that can disrupt the endosomal membrane, or the use of pH-sensitive materials that can undergo conformational changes in the acidic environment of endosomes [7].

Controlled Release
For optimal gene-editing efficiency, it is essential to ensure that the CRISPR components are released at the correct time and location. Nanoparticles can be engineered to release their payload in response to specific triggers, such as changes in pH, temperature, or the presence of specific enzymes. This controlled release allows for sustained expression of the CRISPR components, thereby enhancing the effectiveness of gene editing while minimizing off-target effects and toxicity.

CHALLENGES AND LIMITATIONS

Although nanoparticle-mediated CRISPR delivery holds great promise, several challenges remain in translating this approach into clinical applications

Toxicity and Immunogenicity
Despite their many advantages, nanoparticles can sometimes trigger immune responses or cause toxicity, particularly in the liver and kidneys, where many nanoparticles accumulate. The long-term effects of nanoparticles on the immune system and organs remain a major concern for their use in human therapeutics [8].

Targeting Specificity
Achieving precise targeting of nanoparticles to specific cell types remains a significant challenge. While surface modifications can improve targeting, it is still difficult to achieve high specificity and avoid off-target delivery. Enhancing the targeting efficiency of nanoparticles, especially for tissues or organs that are difficult to reach, remains a key area of research [9].

Manufacturing and Scale-Up
The large-scale production of nanoparticles that meet the necessary quality control and regulatory standards for clinical use remains a significant challenge. Developing scalable methods for synthesizing nanoparticles with consistent properties and controlled delivery efficiency is essential for advancing nanoparticle-based CRISPR therapies.

Regulatory Approval
The approval of nanoparticle-based CRISPR therapies by regulatory bodies is a complex process that requires extensive testing for safety, efficacy, and manufacturing quality. Regulatory challenges, including concerns about potential long-term effects and immune responses, need to be addressed before these therapies can reach the clinic.

FUTURE DIRECTIONS

The future of nanoparticle-enhanced CRISPR delivery lies in improving the biocompatibility, targeting specificity, and scalability of nanoparticles, as well as reducing their toxicity and immunogenicity. Key areas for future research include

Biodegradable Nanoparticles
Developing biodegradable nanoparticles that degrade into non-toxic by-products will be crucial for reducing the long-term risks associated with nanoparticle delivery systems. Biodegradable polymers, such as PLGA and chitosan, are promising candidates for this purpose.

Multi-Functional Nanoparticles
Designing multi-functional nanoparticles that can simultaneously deliver CRISPR components, track their delivery with imaging agents, and monitor gene-editing outcomes will be a game-changer in both personalized and precision medicine.

Exosome-Based Nanoparticles
Exosomes, naturally occurring nanoscale vesicles, are emerging as an alternative to synthetic nanoparticles for gene delivery. Due to their natural origin, exosomes are less likely to provoke an immune response, and they can be engineered to carry CRISPR components for targeted delivery [10].

CONCLUSION

Nanoparticle-enhanced gene delivery is a promising approach for overcoming the barriers associated with CRISPR-based therapeutics. By improving the efficiency, specificity, and safety of CRISPR delivery, nanoparticles hold the potential to transform gene-editing therapies and make them viable for treating a wide range of genetic diseases. Continued advancements in nanoparticle design, targeted delivery systems, and scalable manufacturing will be essential for the successful clinical translation of nanoparticle-enhanced CRISPR therapies.

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