Synthetic mRNAs for Gene Editing

Gene Editing mRNAs for in vitro transfection and in vivo delivery by injection

Footprint-free gene editing without limitations

Nucleus Biotech presents Levostar´s premade mRNAs featuring enhanced stability, prolonged protein expression, and scalable manufacturing capacities.

Levostar´s mRNAs encoding CRISPR Cas enzymes, recombinases, as well as transposases enable various gene editing approaches such as CRISPR Knock Out, Base Editing, Prime Editing, as well as Recombinase/Transposase – supported integrations, all in a footprint-free manner.

• No Size Limitations

mRNA does not have the size limitations associated with AAVs or other viral particles, which allows high expression of large and multi-domain gene-editing enzymes, increasing gene editing efficiency and broadening the scope of gene editing technologies

• Direct Applications in Animal Studies

In vitro gene editing results can be extended to animal studies facilitated by in vivo mRNA injections. This leads to a more comprehensive understanding of your gene-editing outcomes and supports your translational research efforts.

• Ease of Use

mRNAs – especially when produced by Levostar´s streamlined manufacturing procedures – are much easier to apply than RNPs or AAVs and results are much more reproducible

Levostar mRNAs are designed for optimal expression results:

• Enhanced polyA(+) Stability – for preventing RNA degradation

• Cap1 Structure – for optimal ribosome recruiting

• Carefully selected UTR Regions – to enhance translation initiation

N1-Methylpseudouridine Modification

For enhanced mRNA stability and decreased anti-mRNA immune response Levostar´s catalog mRNAs are in vitro transcribed using N1-Methylpseudouridine (N1-Me-Ψ, N1mpU) by default. If you prefer unmodified mRNA or you require a different modification, please contact us.

monoPlasmid DNA Technology

Levostar mRNAs are manufactured by their proprietary monoPlasmid Technology enabling high In Vitro Transcription (IVT) yields and outstanding polyA(+) stability:

monoPlasmid: IVT Plasmids with high monomer ratio

Due to the presence of the polyA tail, mRNA plasmids are highly prone to inter-plasmid recombination, resulting in the formation of large plasmids as dimers or multimers. Multimer formation significantly reduces plasmid yield. LevoStar´s monoPlasmid platform incorporates plasmid elements as well as a proprietary bacterial strain, together preventing multimer formation, which ensures a plasmid monomer supercoiling rate of over 95% while also substantially increasing plasmid yield.

Common mRNA plasmids show a high ratio of multimers

monoPlasmid mRNA plasmids show a high ratio of monomers

monoPlasmid: IVT Plasmids with stabilized polyA(+) tails

Due to its low free energy, the polyA sequence in mRNA plasmids is prone to breakage within E. coli, leading to partial or complete loss of the polyA tail during plasmid repair. Such polyA-deficient mRNA plasmids cannot be used to produce functional mRNA. Levostar´s monoPlasmid platform incorporates a specialized sequence that significantly reduces the likelihood of such loss.

Sanger Sequencing demonstrates partial loss of the polyA tail in common mRNA plasmids

Sanger Sequencing demonstrates complete polyA tail presence in monoPlasmid mRNA plasmids

Quality Standards: All premade mRNAs are tested for high purity, low endotoxin,  low residual dsRNA & plasmid template DNA, high capping efficiency, and proper polyA(+)-length.

We are pleased to offer premade mRNAs encoding CRISPR Cas enzymes, recombinases, and transposases.

In case you can´t find your mRNA of interest, you can use our mRNA synthesis services.

We are also offering Levostar´s NeoLNP product line for easy and highly efficient mRNA encapsulation and LNP-based in vitro mRNA transfection as well as in vivo mRNA delivery by injection.