Seth Cheetham

The design, manufacture and LNP formulation of mRNA for research use Nature Protocols - This protocol describes simple, small-scale laboratory production of mRNA for research and preclinical use. This includes sequence design, DNA template production, mRNA synthesis...

To accelerate #mRNA vaccine and therapy research, we're excited to share our end-to-end protocol for mRNA design, synthesis and encapsulation in @natprot.nature.com.

Protocol development led by @laura-leighton.bsky.social, Natasha Chaudhary and Tim Mercer.@basemrna.bsky.social

rdcu.be/eqiF1

Today, a huge milestone for personalised #mRNA therapeutics for inherited disease! A baby with an ultra-rare mitochondrial metabolic disease was treated at UPenn with an mRNA-encoded gene-corrector. The mRNA was made to treat a single individual in just 7 months!

www.nejm.org/doi/full/10....

Crucially, this approach doesn't require a new LNP, it works with FDA-approved formulations. We hope that this approach will enable targeted mRNA therapeutics for diverse diseases. Work led by Bettina Dietmair, James Humphries and Chris Howard. @basemrna.bsky.social

The technology is programmable and works with other targeting arms including anti-PSMA (highly expressed on prostate cancer cells) bispecifics. 6/

Bispecifics can be used by combining with the mRNA-LNP (pre-mixing) or the bispecific can be administered first (pre-targeting). 5/

As proof of concept we used anti-EGFR bispecifics to deliver mRNA to mouse breast cancer xenografts. The mRNA-payload (luciferase) is very efficiently delivered in vivo. 4/

To overcome this obstacle we developed bispecific antibodies that bind to PEG on the surface of LNPs and to cell-surface markers. 3/

Getting a high dose of mRNA to target tissues is one of the toughest problems for broad use of mRNA-LNP therapies. When administered systemically mRNA-LNPs go the liver with little delivery to other tissues. 2/

Targeted mRNA delivery with bispecific antibodies that tether LNPs to cell surface markers Efficient delivery of mRNA-lipid nanoparticles (LNPs) to specific cell types remains a major challenge for mRNA therapeutics. Conventional targeting a…

Systemic mRNA-LNP delivery beyond the liver? Check out our recent paper demonstrating cell-specific mRNA-LNP delivery using PEG- and target cell-specific antibodies. This approach can customised to any target cell-type with off-the-shelf mRNA-LNPs. 1/
sciencedirect.com/science/arti...

Exited to share our latest preprint! mRNA-LNPs are efficiently delivered to healthy livers, but what about liver tumours? Our postdoc @laura-leighton.bsky.social found liver tumours efficiently take up mRNA, enabling next generation mRNA-LNP cancer therapeutics.

www.biorxiv.org/content/10.1...

Developmental biology research leads to fundamental discoveries for biomedical research. RNA Vaccines. Lane et al., 1971 PMID: 5169490. Images by Brandy Walker & Amaya Moss @rkmille . @socdevbio.bsky.social 3/12

Happy to sure our protocol if helpful, just send me a DM

We use q5 which has a very low error rate. We've done a lot of ONTseq of plasmid and pcr templates and don't see any increase in error rate, but we see way better polyA integrity.

How much mRNA are you producing? We routinely use PCR to generate templates for up to 1-20mg productions. I know people that have done larger batches too.

There are some tweaks like segmented polyA tails that somewhat reduce deletions, but they're still very frequent. Worse, they are sometimes subclonal so sanger sequencing doesn't even detect the deletion. The field definitely needs better synthetic biology solutions.
doi.org/10.1261/rna....

Calling on #ONT #Nanopore #sequencing experts: when sequencing through a stretch of 120nt dA using a DNA plasmid kit (outsourced in our case), what should one expect as average length in the output? The loss we are seeing is a bit extreme #RNAsky

Could be real though, polyA deletions in plasmids are really common and one of the reasons we switched to PCR-based templates.

Radioprotection of healthy tissue via nanoparticle-delivered mRNA encoding for a damage-suppressor protein found in tardigrades - Nature Biomedical Engineering Radiation-therapy-induced DNA damage in oral and rectal epithelial cells can be reduced via the nanoparticle-mediated local delivery of messenger RNA encoding for a damage-suppressor protein found in ...

#mRNA meets Tardigrades! Delivery of an mRNA encoding a protein from the famously tough animal can protect healthy tissue from damaging radiation. Very creative! #RNAsky

www.nature.com/articles/s41...

@helengunter-mrna.bsky.social is the expert at this.

Not the best weather forecast for Brisbane 😬

Preach!

You might be a bit overqualified Scott ;)

Interested in undertaking a PhD in the transformative field #mRNA cancer vaccines? I have an earmarked #PhD scholarship available for an outstanding domestic student to join our team at UQ and @basemrna.bsky.social. If this sounds like you, get in touch.

study.uq.edu.au/study-option...

CleanCap M6 inhibits decapping of exogenously delivered IVT mRNA In this manuscript, Mandell and colleagues present CleanCap M6, a next generation 5’ cap analog that outperforms existing industry standards in live mice. Mandell and colleagues found that this cap ex...

New #mRNA cap structures get you more bang (protein) for your buck! Nice new research from @jmcoller.bsky.social in collaboration with Trilink shows the CleanCap M6 analogue inhibits decapping and substantially increases protein expression in vivo.

www.cell.com/molecular-th...

Nanocarrier imaging at single-cell resolution across entire mouse bodies with deep learning - Nature Biotechnology An integrated experimental and deep learning pipeline reveals cell-level targeting of nanocarriers in whole bodies.

Wow this is super cool! Single-cell resolution imaging of #mRNA-LNP biodistribution in cleared mouse bodies.

www.nature.com/articles/s41...

UQ and Vaxxas win prize to advance patch-based mRNA COVID-19 vaccine UQ’s BASE facility and biotechnology company Vaxxas have been announced as stage winners of a prestigious global prize to accelerate a patch-based mRNA vaccine for COVID-19.

Exciting news! @basemrna.bsky.social and Vaxxas have been awarded the $2M USD Biomedical Advanced Research and Development Authority (BARDA) Patch Forward Concept Stage Prize to develop thermostable needle-free mRNA vaccines.
www.uq.edu.au/news/article...

Enhancement of anti‐sarcoma immunity by NK cells engineered with mRNA for expression of a EphA2‐targeted CAR Human NK cells can be efficiently transfected with mRNA to safely express an EphA2-CAR without genomic alterations. EphA2-targeted CAR-NK cells exhibit enhanced cytotoxicity against paediatric sarco...

#mRNA meets cell therapies! It was great to collaborate with Fernando Guimaraes's team to develop mRNA CARs and delivered them to NK cells. NKs transfected with EphA2 CAR mRNA elicited a strong anti-tumour response in animal sarcoma models.

onlinelibrary.wiley.com/doi/10.1002/...

Out of office

Placenta-tropic VEGF mRNA lipid nanoparticles ameliorate murine pre-eclampsia - Nature A platform for mRNA lipid nanoparticle delivery to the placenta to treat pre-eclampsia is shown to improve fetal and maternal health in mice and has potential clinical applications in obstetric d...

Awesome work from Michael Mitchell's lab at UPenn in @natureportfolio.bsky.social identifies novel LNPs that can deliver #mRNA to the placenta. Using the placenta-targeting LNPs to deliver VEGF mRNA improves pre-eclampsia in animal models.

www.nature.com/articles/s41...

YouTube video by TEDx Talks How personalized mRNA vaccines could revolutionize cancer treatment | Seth Cheetham | TEDxUQ

Earlier this year I had the opportunity to present a TEDx talk on the transformative potential of personalised #mRNA vaccines for cancer. The recording is out now:

www.youtube.com/watch?v=BnSO...

#RNAsky #RNAtx @basemrna.bsky.social