RiboPro offers custom lncRNA synthesis, so you can get started with your experiment as quickly as possible, without hassle!
No need to worry about RNAse contamination during production, no need to setup complex quality control, no need to invest in the equipment and materials; we take care of it all.
Use the lncRNAssembler™ below to design your lncRNA, instantly see the price and simply add the designed sequence to your cart to order. For a typical sequence, you will receive your order in 2-3 weeks.
Do you want to know more before making a decision? Scroll down for additional info and reviews at the bottom of this page.
lncRNAssembler™ – quick and easy ordering of your lncRNA
The performance of a long non-coding RNA, to be added to cells via standard transfection methods, differs from messenger RNA; there is no need for protein production. However, similar to in vitro transcribed (IVT) messenger RNA, IVT long non-coding RNA can trigger unwanted innate immunogenicity, effecting cell-wide transcription levels, potentially affecting experimental readout. These potential side-effects greatly depends on the design of the sequence, the quality of the synthesis, chemical modifications of specific bases and the purity of the end-product.
For the best performance, we recommend the following combination of features to be selected for your custom lncRNA (if lncRNA is endogenously synthesized by RNApol II and bears a cap and A-tail):
Cap1 – 95%.
120-150nt A-tail.
dsRNA reduction.
optional pre-folding by rapid or slow cooling after heating.
If you like to understand why these settings lead to the best performance, read more below.
Synthesizing a high-performance long non-coding RNA requires optimization all through-out the multi-step synthesis, including specialized purification.
First of all, during the enzymatic production process, dsRNA can be formed. dsRNA is one of the main triggers of innate immunity, primarily via TLR3 (endosomal recognition during transfection), OAS1, PKR and MDA5 (cytoplasmic sensors). RiboPro’s mRNA synthesis process results in the lowest amount of dsRNA possible. The method used reduces the yield of single synthesis, so additional costs (limited amount) do apply compared to the standard method, because we increase the scale in order to obtain the guaranteed yield. Without dsRNA reduction, protein expression is typically 10-fold lower than with dsRNA reduction.
Secondly, currently no supplier can promise that 100% of all mRNA molecules will be capped. When uncapped, the 5’end of the lncRNA contains a 5′ triphosphate, which can be a ligand for the cytoplasmic innate immune sensor RIG-I. RiboPro offers 2 capping grades (~80% and 95%+). If selecting the lower grade (cheaper), additional phosphatase treatment may be required to reduce RIG-I activation.
Third, if a cap is present, but no additional methylation of the first nucleotide (Cap0), then IFIT1 may be activated, leading to around 30-50% reduction in activity in some experiments. RiboPro offers optional methylation of the first nucleotide (Cap1).
Finally, removal of template DNA, salts and small RNA species (formed during abortive cycling of the RNA polymerase) must be carefully removed to avoid interfering with uptake and interpretation of results.
Next to an optimized production process, RiboPro also has also developed a series of test for quality control, offering you peace of mind about the identity, purity and modification status of your order.
Do you have additional questions? Please contact us below or later via the contact page. We are happy to help!
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