Extend protein expression with Poly(A)+ services for custom mRNA
A poly(A) tail modification on mRNA may serve several important functions that benefit the molecule's stability and functionality to help enable next-generation RNA medicines.
TriLink's proprietary novel poly(A) tail modification increases overall protein expression and extends the duration of protein expression. The poly(A) tail modification is now available through TriLink's custom mRNA synthesis services.
TriLink’s poly(A) tail modification increases the protein expression and duration
The TriLink’s poly(A) tail modification is enabled via our Poly(A)+ services. TriLink’s poly(A) tail modification is compatible with various caps, base modifications, construct sizes, and poly(A) tail lengths.
TriLink’s poly(A) tail modification increases eGFP protein expression and duration in HEK293T cells.
HEK293T cells were transfected with CleanCap® M6 capped mRNA encoding eGFP with or without TriLink’s poly(A) modification. CleanCap® M6 capped eGFP protein fluorescence was monitored over time by Incucyte.
Mice injected with FLuc mRNA with a 120A tail with TriLink’s poly(A) modification show increased duration of protein expression.
Mice were injected with PBS (top rows), or Fluc mRNA:LNPs for whole body luminescent imaging at 12, 48, and 72 hours. For both CleanCap® AG 3’OMe (left panel) and CleanCap® M6 (right panel) capped Fluc TriLink’s poly(A) tail modification (bottom rows) resulted in higher expression signal at later timepoints.
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Leverage TriLink's Poly(A)+ services to customize your mRNA with our new poly(A) tail modification via our mRNAbuilder® Tool
Let’s discuss your mRNA program needs
If you are ready to submit a custom RNA synthesis request with the poly(A) tail modification, use the mRNAbuilder® tool.
For all other inquiries, tell us more about your specific program needs so we can tailor our initial conversation with you. We look forward to connecting with you.
Poly A tail Modification form (CDMO)
Frequently asked questions (FAQs) about TriLink’s Poly(A)+ services
General
TriLink’s proprietary poly(A) modification is a small chemical moiety linked to the 3’-end of the poly(A) tail of mRNA after the in vitro transcription reaction. There are no other changes to the mRNA.
No, the modification is not template-directed and occurs post-IVT. In this case, there is no need to change your DNA template, and there is no impact on the mRNA coding sequence.
Yes, we have tested our proprietary poly(A) tail modification with different poly(A) tail lengths including 40A, 60A, 80A, 100A, and 120A. We have successfully added our Poly(A) tail modification to various mRNAs, such as eGFP, Fluc, mCherry, beta-gal, and Cas9 mRNA. These constructs range from 0.9 to 4.5 kb in length.
TriLink’s poly(A) tail modification should be compatible with commonly used delivery vehicles. We have used it with common transfection reagents and LNPs, including LipofectamineTM MessengerMAXTM reagents, and ALC-0315, DSPC, Cholesterol, and DMG-PEG 2000: Pfizer/Acuitas Lipids.
Quality
No, we have data showing that the mRNA is of comparable integrity to regular oligo-dT-purified mRNA.
We have an analytical poly(A) modification efficiency assay to verify the incorporation of TriLink’s poly(A) tail modification. We verify the poly(A) modification by analytical RP-HPLC. The Analytical Report for this service will include results for each construct.
Function
The precise mechanism of how TriLink’s Poly(A) modification increases the protein expression and duration is currently under investigation; however, we hypothesize it prevents 3’-exonucleosase cleavage as one of the mechanisms.
We have not observed meaningful differences in the immunogenicity of tail-modified mRNA from non-tail-modified mRNA in our in vitro and in vivo studies. Please see the technical note to learn more.
Access
It is currently available through our Discovery (RUO) mRNA synthesis services. You can request a quote or order through our mRNABuilderâ„¢ configuration tool.
Yes, we can receive your material as customer-supplied material (CSM) and then perform the tail modification. We require purified mRNA along with the following information: mRNA length, tail length, concentration, and target quantity of modified mRNA. Please email sales@trilinkbiotech.com for a quote.
Our research use only (RUO) terms and conditions cover research use of TriLink’s proprietary poly(A) tail modification. If you have additional questions, reach out sales@trilinkbiotech.com. Should you require expanded use rights, a separate agreement may be negotiated between you and TriLink.