A deep dive into the world of gene therapy
Gene therapy is promised to be the answer for the treatment or prevention of many genetic diseases, to address diverse clinical applications, from cancer to neurological disorders and inherited diseases, and can potentially provide long-lasting or even permanent treatment solutions.
Their mechanism of action is based on the delivery of genetic material, and the two primary types of nucleic acids used in gene therapy are:
- DNA (Deoxyribonucleic Acid): DNA is the molecule that encodes genetic information in living organisms. In gene therapy, DNA can be delivered into a patient’s cells to replace or supplement a defective gene, or to introduce therapeutic genes. This is often done using viral (modified viruses) or synthetic vectors that can deliver the therapeutic DNA into target cells.
- RNA (Ribonucleic Acid): RNA plays a key role in the process of protein synthesis based on the information encoded in DNA. There are different types of RNA used in gene therapy: mRNA for protein production, siRNA to silence genes, miRNA for gene expression regulation, and sgRNA CRISPR for precise gene editing, among others.
The potential of mRNA
The ability to modulate gene expression upon delivery of mRNA has already been proven to be effective for the development of vaccines and other applications. They are especially effective for producing temporary therapeutic effects, responding to emerging infectious diseases like in COVID-19 vaccines, as well as to unmet clinical needs in prevalent and rare diseases. mRNA has several advantages over other gene therapies:
- Higher transfection efficiency.
- Higher safety, thanks to the inability to integrate into the genome, and thus, less mutagenesis risk.
- Rapid development potential thanks to the ease for production and modification of the sequence as desired, for personalized treatments.
However, when we speak about converting these molecules into medicines, there are two main disadvantages all nucleic acids have. First, they are large negatively charged biopolymers which are not capable of crossing lipid bilayers. Furthermore, their naked administration can promote immune reactions. Like so, the development of safe and effective delivery vehicles is crucial for the translation to the preclinical and clinical development.
mRNA delivery methods available in gene therapies
The delivery of RNA molecules into cells is a crucial aspect of various biomedical and therapeutic applications, ranging from gene silencing to vaccine development. There are several methods used to deliver RNA molecules into cells, each with its own advantages and limitations. Here are some of the main types of RNA delivery methods:
- Mechanical methods, such as electroporation and sonoporation, create temporary pores in cell membranes that enable RNA entry.
- Chemical methods, relying in the formation of complexes with cationic lipids or polymers that favor interactions with cell membranes and RNA entry.
- Viral vectors, which can be engineered to carry RNA molecules and deliver them to target cells.
- Non-viral delivery systems, such as Lipid Nanoparticles (LNPs), which are a type of lipid-based nanoparticle that have been used for delivering RNA-based therapeutics, including mRNA vaccines, and have reached a clinical setting. The use of LNPs and other lipid nanosystems, as DIVERSA delivery technology, has been extensively reviewed by Taina-Gonzalez and De La Fuente M.
Mechanical and chemical methods typically harm cells and show toxicity, while cannot be applied in an in vivo setting. Talking about viral delivery, these are effective in delivering the genetic material to the nucleus of the cells and well as helping integrate the DNA in the genome of the host. However, despite the success of AAV-based drugs, around 50% of the patients are excluded from treatment because of preexisting immunity to viral capsids. In this regard, non-viral delivery systems can provide a solution towards the clinical approval of more gene therapies.
At DIVERSA we specialize in lipid-based delivery systems, using our safe and patented technology and different strategies to associate nucleic acids.
DIVERSA innovative lipid nanoemulsions for mRNA applications
Whether you’re working with small or large mRNA sizes, our formulation accommodates them all, opening a broad spectrum of research possibilities. We have an in vitro proof of concept for both luciferase (Fluc mRNA – 2000 bp) and fluorescence (eGFP mRNA – 1000 bp) mRNA. On a separate note, the technology has also been successfully employed for smaller RNAs and bigger DNAs.
Characterization of nanoemulsions: Size (nm) and homogeneity represented as Polydispersity index (PDI)
Our reagent operates without the need for adeno-associated viruses (AAV), ensuring a safe and efficient mRNA delivery.
We take cell integrity seriously. Our reagent is bio-inspired, ensuring optimal biocompatibility, opposite to what observed with transfection reagents. Our formulation does not rely on mechanical and chemical methods that increase toxicity.
HEPG2 cells after 48h incubation with DIVERSA mRNA Delivery Reagent loaded with FLuc-mRNA treatment compared to Messenger MAX
Our system ensures a high intracellular delivery, speeding up your experiments and getting you to results faster. We provide comparable results in vitro and in vivo to a composition that has reached the clinic.
This highlights the value of DIVERSA’s formulation, standing out from reagents based on chemical method that do not allow an easy translation from in vitro to in vivo.
In vitro and in vivo transfection with FLuc mRNA associated to DIVERSA’s technology (DIV:FLuc mRNA) and to a control formulation (FLuc mRNA, based on commercialized Onpattro® composition).
Example of a successful case: THERAPEUTIC EFFICACY OF IL-12 mRNA
At DIVERSA we maintain a constant R&D activity and we partner with highly reputated scientists.
In the framework of Laura Taina’s Industrial PhD, we have proved the potential of our technology to deliver therapeutic mRNA encoding an interleukin with anticancer activity before and interfere tumor growth. This work has recently been presented in the biggest event in drug delivery and nanomedicine, the Controlled Release Society Annual Meeting, Laura Taina being awardee of a competitive travel grant to attend.
Do you have a potential therapeutic nucleic acid but you lack an effective delivery strategy?
Try our mRNA DELIVERY REAGENTS:
The best way to have a first contact with our technology is without a doubt trying our ready-to-use Delivery Reagents, specifically designed for this purpose and with a user friendly methodology. However, this does not mean that we can’t help you with any questions that you may have regarding your peptide. Please, don’t hesitate to contact us to order your DIVERSA Delivery Reagent or if you need further guidance.
Establish a CO-DEVELOPMENT AGREEMENT:
We understand that you may wish to have a more personalized approach, and for this reason, we have created the Co-Development Agreements. In this case, we would build up a specific project for your specific project.
You can contact us or write an email to email@example.com. We are looking forward to helping you with your molecules!
DIVERSA technologies aims to provide delivery solutions through the design of new and safe vehicles, easy to produce and versatile, which can be adapted to different types of therapeutic molecules (both biomolecules and drugs) to facilitate their release, especially when their target is found at the intracellular level and access is limited. DIVERSA is focused on promoting the transfer to the clinic of new therapeutic molecules with high potential and high added value, based on a patented technology.