Continuous Nanoparticle Manufacturing For Gene Editing Delivery

Gene editing technologies such as CRISPR/Cas9 are redefining approaches to treating genetic disease by allowing precise modifications to DNA. Their effectiveness, however, depends largely on how the editing machinery is delivered into cells. Messenger RNA encapsulated in lipid nanoparticles (mRNA-LNPs) has become a leading delivery strategy. LNPs protect RNA molecules, promote cellular uptake, and enable transient expression of editing components.

While mRNA-LNPs show biological advantages, translation into clinical use also requires manufacturing methods that can reliably produce nanoparticles with consistent properties. The reproducibility, scalability, and monitoring of production are therefore central to enabling further applications. DIANT Pharma has developed a continuous manufacturing system intended to address these requirements.

Why mRNA-LNPs Are Used in Gene Editing

For CRISPR/Cas9 applications, mRNA-LNPs offer several practical advantages. Once inside a cell, mRNA directs production of the Cas9 nuclease and guides RNAs needed for DNA targeting. Because mRNA naturally degrades, expression of Cas9 is temporary, which helps reduce the risk of unwanted off-target activity. RNA also avoids permanent integration into the genome, lowering the likelihood of insertional mutagenesis.

The clinical familiarity of LNPs, established through their use in mRNA vaccines, further supports interest in this approach. Key features include:

• Transient activity – Cas9 expression is limited in duration.
• Non-integration – RNA avoids permanent genetic changes.
• Existing validation – LNPs are already in clinical use for RNA delivery.

These properties make mRNA-LNPs a practical platform, provided that manufacturing processes can consistently produce particles with controlled quality attributes.

The Manufacturing Challenge

Nanoparticle performance depends on physical and chemical properties such as particle size, charge, and encapsulation efficiency. Variations in these parameters can influence biodistribution, uptake, and safety. Batch-based production methods often introduce variability, require multiple manual steps, and can be difficult to scale without re-optimization.

For gene editing applications, where precision and reproducibility are critical, these challenges highlight the need for alternative manufacturing strategies.

DIANT Pharma’s Continuous Manufacturing Platform

DIANT Pharma has developed a continuous, closed-loop nanoparticle manufacturing system that integrates particle formation, purification, concentration, and monitoring. The design is intended to improve reproducibility, reduce manual interventions, and support process scalability.

Notable features include:

• Controlled particle formation – Jet mixing produces LNPs with narrow size distributions, which is important for predictable biodistribution.
• Single-pass tangential flow filtration – Impurities are removed and buffers exchanged in a single pass, minimizing product stress.
• Integrated downstream steps – Concentration and sterile filtration are built in, reducing the need for multiple systems.
• Real-time monitoring – Analytical tools provide immediate feedback on process parameters, supporting in-process control.

This approach allows the same system to be applied from laboratory research to larger-scale production, which may reduce the need for re-engineering during scale-up.

Potential for Tailored Delivery

Adjusting nanoparticle properties such as lipid composition, particle size, and surface chemistry can influence biodistribution. A system capable of fine control over these parameters can be used to explore targeted delivery of CRISPR components to specific organs.

Potential areas of application under investigation include:

• Liver – a frequent target for nanoparticle delivery in metabolic and storage disorders.
• Lungs – relevant for genetic respiratory diseases such as cystic fibrosis.
• Heart and skeletal muscle – associated with certain inherited cardiomyopathies and myopathies.
• Central nervous system – a challenging area for delivery but an active area of research.

Regulatory and Development Considerations

Continuous manufacturing offers potential advantages in meeting regulatory expectations for consistency and quality. Built-in monitoring tools support process analytical technologies (PAT), which emphasize quality assurance during production rather than relying solely on end-product testing.

Because the platform is designed to be scalable, formulations optimized in early-stage studies can be manufactured at larger volumes without major redesign, potentially streamlining development timelines.

Delivery is a central challenge in the development of CRISPR/Cas9-based therapies. Biological requirements point toward transient, efficient, and safe systems such as mRNA-LNPs, while development requirements emphasize reproducibility, scalability, and regulatory alignment.

DIANT Pharma’s continuous manufacturing technology is designed to address these needs by enabling controlled, consistent production of mRNA-LNPs. While additional clinical validation is required, the approach provides a structured framework for advancing nanoparticle delivery in gene editing applications.

Source: https://www.bioprocessonline.com/ecommcenter/diant_pharma