Gene Therapy

Gene Therapy modulates gene expression to provide root-cause treatment by targeting the aberrant genes / gene networks responsible


Precision NanoSystems’ Genetic Medicine Toolkit provides essential technologies to facilitate development of Gene Therapies that offer new avenues to treat some of the most challenging diseases.

Gene Therapy Toolkit

PNI Genetic Medicine Technologies

Non-viral Gene Therapy is a viable therapeutic approach for diseases arising from a loss-of-function mutation by enabling protein replacement therapy. The Genetic Medicine Toolkit was used to demonstrate proof-of-concept (PoC) for development of a protein replacement therapy in a model of anemia. The drug product consisted of an mRNA encoding erythropoietin encapsulated in a GenVoy lipid nanoparticle (LNP) using the NanoAssemblr manufacturing platform. The Non-viral Gene Therapy approach has several key advantages:


Decreased complexity: no cell culture required to produce the drug product, simplifying R&D and manufacturing.

Increased Modularity: mRNA can be designed to specifically express any protein, and optimized LNP formulations and drug product manufacturing can be leveraged to streamline development of future genetic medicines.

Non-Viral Gene Therapy mediates Erythropoietin Production Reversing Disease Phenotype in an Anaemia Model



First, mRNA designed to express human Erythropoietin (EPO) was encapsulated in GenVoy Lipid Nanoparticles (LNP) using the NanoAssemblr manufacturing platform

Second, following intravenous administration, GenVoy LNP delivered mRNA to hepatocytes in the liver which expressed and secreted EPO into the bloodstream to stimulate red blood cell production
Lastly, the Epo mRNA-LNP mediated increased red blood cell production in the anaemia model. These results were consistent as the Non   Viral Gene Therapy production was scaled using the NanoAssemblr manufacturing platform

Watch the webinar presented by Samuel Clarke, PhD – Director of R&D



How to Develop a Gene Therapy


The development process for genetic medicines is similar to other therapeutic modalities and can be illustrated using a Framework highlighting the key stages and milestones needed to progress a genetic medicine from idea to new disease treatment. The example below demonstrates how the elements of the Genetic Medicine Toolkit are designed to accelerate Non-Viral Gene Therapy development by enabling the work required in each stage of the Framework.

PNI’s Genetic Medicine Framework and Toolkit Guides and Enables Development of Non-Viral Gene Therapy


GenVoy ILM-Lipid Library V2-01

The Target Product Profile (TPP) is used to identify the ideal profile of a drug product for treatment of a target disease. While the TPP for a given Gene Therapy depends on characteristics such as the disease, the drug target, and the market landscape, an example of a simplified profile is shown below for illustrative purposes:


Vaccine Characteristics

General Requirements


Once every 2-4 weeks.

Treatment Duration


Route of Administration


Product Stability and Storage

Long term storage at -20℃ or higher.


Single dose presentation; Liquid suspension.

Maximum parenteral dose volume: 20mL.

Disease Target-8


1. Target Selection

Targets are often identified through bioinformatic approaches utilizing genomic information associated with the disease. Disease states may be targeted by knocking down genes using siRNA or expressing genes through mRNA. Approaches using gene editing such as CRISPR/Cas9 will result in more durable, less frequent treatments. Targets identified in silico are validated in vitro using relevant cell culture, and in vivo using phenotypic models.



2. Vector Selection

For gene knockdown, a double stranded siRNA can mimic microRNA and mark a target for degradation through the RNA interference (RNAi) pathway [1].

To express an exogenous gene, mRNA is preferred for transient expression, ease of delivery by non-viral carriers and low risk of genome integration. For protein replacement therapies, mRNA featuring a cap1 structure and mammalian base modifications are preferred to minimize immune reaction and maximize half-life. [2,3]

Both siRNA and mRNA can be manufactured by cell-free enzymatic synthesis that reduces the complexity of manufacturing and greatly accelerates development of RNA therapeutics. [4,5]

Self-Amplifying RNA


mRNA encoding human erythropoietin, synthesized with cap1 structure and base-modified uridines (5-methody uridine) and delivered by LNP were found to increase red blood cell production in mice. This provides proof of concept for mRNA-LNP based protein replacement therapy.




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LNP icon-01

3. Delivery and Formulation Technology

LNP are the most advanced non-viral gene delivery systems. In 2018, the FDA approved ONPATTRO® (patisiran) an siRNA-LNP gene therapy for the treatment of polyneuropathy precipitated by hATTR amyloidosis. More recently, mRNA-LNP COVID-19 vaccines have been authorized for clinical use in several countries. PNI’s LNP technology (GenVoy) uses ionizable cationic lipid-based formulations similar to those employed in approved products. 

Unlike viral vectors, GenVoy RNA-LNP are produced using a cell-free, scalable continuous flow process that can streamline and accelerate development of non-viral Gene Therapy. 

PNI’s LNP Delivery Technology


Serum protein levels were equivalent  6h post-injection

Erythrocyte levels were elevated equivalently 7 Days post-treatment

Erythrocyte Levels

Epo mRNA-LNP prepared using GenVoy demonstrated equivalent activity in vivo to Epo mRNA-LNP prepared using a MC3 lipid-based formulation. MC3 is the ionizable cationic lipid used in ONPATTRO.



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4. Manufacturing

The NanoAssemblr® platform uses microfluidic mixing to manufacture RNA-LNP. The proprietary NxGen microfluidic architecture has been designed and optimized to exquisitely control self-assembly of the RNA-LNP to ensure reproducible manufacture of high quality drug products.


The NanoAssemblr platform comprises Spark, Ignite, Blaze and GMP Systems, which are designed to support development of non-viral gene therapy from discovery to commercial production. Transferring RNA-LNP production between Systems during development is simplified because they all use the NxGen microfluidic mixing technology. This concept is illustrated in the figure below

NanoAssemblr™ Technology: Scalable RNA-LNP Manufacturing
Epo-Encoded mRNA-LNP1
Epo-Encoded mRNA-LNP 2

Epo-encoded mRNA-LNP had similar size (~70 nm), polydispersity (<0.1) and encapsulation efficiency (>90%) across NanoAssemblr instruments, demonstrating streamlined scale up of LNP production across all stages of genetic medicine development

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End-to-end Gene Therapy Development Solutions

Precision NanoSystems is committed to providing clients with flexible access to technology and expertise to accelerate development of new gene therapies from idea to commercial product.

Genetic Medicine Development Framework


The Framework provides a roadmap for genetic medicine development, including non-viral gene therapy

Genetic Medicine Toolkit


Essential technologies to facilitate development of non-viral gene therapy




Kick-start development of non-viral genetic medicines with focused virtual classroom and hands-on training.

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2.      Karikó, K., Buckstein, M., Ni, H., & Weissman, D. (2005). Suppression of RNA Recognition by Toll-like Receptors: The Impact of Nucleoside Modification and the Evolutionary Origin of RNA. Immunity, 23(2), 165–175. https://doi.org/10.1016/j.immuni.2005.06.008
4.      BioLabs, N. (2018). RNA Synthesis FROM TEMPLATE TO TRANSCRIPT. https://www.neb.com/-/media/nebus/files/brochures/rna-brochure.pdf?rev=a319b91c67d845458b22d29ee00fa53a

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