[On-Demand Webinar] [Case Study] Tracking Tools for mRNA Therapeutic Research: A practical example using a 3D colorectal cancer model

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What is this webinar about?

mRNA vaccines proved their value during the Covid-19 pandemic and are now paving the way for therapies against other diseases, notably cancer. High-quality RNA production and delivery are therefore crucial for oncology research.

In this case study, Erica Cirri, PhD outlines the critical steps: designing and producing an mRNA, creating an in vitro 3D colorectal cancer (CRC) model, and successfully encapsulating the mRNA in lipid nanoparticles (LNPs) for delivery into HCT116 spheroids.

Fluorescently labelled molecules enabled tracking of both the mRNA and LNPs, confirming their incorporation and the expression of the target protein. This preliminary work identified a benchmark LNP formulation for future functional testing of the RNA-based therapy.

Webinar summary

mRNA production
DNA template design for mammalian cell expression
mRNA synthesis and modification
LNP formulation
Screening and customisation of LNPs formulations
Production of fluorescently labelled LNPs
3D models
Spheroids generation (colorectal cancer model)
Molecular tracking (protein expression and LNP fate)
Cellular monitoring (cytotoxicity)
Q&A session with Tebubio project managers

Q&A of the live session

Are you able to encapsulate in LNPs molecules other than mRNA?

Yes, we have successfully encapsulated DNA oligoes and saRNA.
Which types of fluorescent lipids do you propose?

We currently propose either phospholipids or PEGylated lipids. If you need something different, our partners can provide custom products.
Can you explain better the role and mechanism for polycistronic mRNA candidate and control to monitor mRNA expression?

A polycistronic mRNA codes for multiple proteins. In the example shown in the webinar, the mRNA transcript codes for the sequence of interest plus eGFP at the 3'. The mRNA is initially translated into a single amino acid sequence; thanks to the presence of an autocleavable peptide (like T2A and P2A) between the proteins, this amino acid sequence autocleaves itself to release each protein separately. This strategy enables monitoring of translation of the protein of interest while maintaining its functional activity unaffected.
Do you have 3D-models to evaluate LNPs for cancer therapies, and offer services on it?

Yes, thanks to our partners, we have access to a large panel of cell lines and primary cells that can be used in service to generate spheroids, organoids or tumoroids.
Could you make these LNP cell type specific?

Some of our partners provide lipids with tropism for liver, lung, or spleen, or more specifically for T cells. Otherwise, it would be necessary to functionalize the LNPs with specific ligands for targeting. Depending on your needs, this might require some development.