Circular RNA（circRNA）is a unique class of non-coding RNA molecules, They are formed through a mechanism called back splicing, The two exons of the precursor mRNA are connected into a closed loop structure during splicing, This avoids the common 5' cap and 3' tail structures found in linear RNA molecules. This circular structure confers high stability on circRNA, Make it less susceptible to degradation by nucleases within the cell. Therefore, they have a longer half-life in cells, which allows them to achieve low-dose and long-lasting effects in practical applications. In addition, circRNA has the advantage of a relatively simple production process because it does not require the modified nucleotides the capping or polyadenylation, making circular RNA a hot spot in drug development.
It has significant advantages in drug development, the details of which are mainly listed in the following:
01 Higher stability
The structure of circRNA makes it resistant to degradation by nucleases, so it has a longer half-life and higher stability in cells, which is crucial for the sustained effect of drugs.
02 Low immunogenicity
Compared to linear RNA, circRNA has lower immunogenicity, which reduces the possibility of immune reactions, making circRNA more suitable as a drug carrier.
03 Efficient protein expression
Research shows that engineered circRNAs can stably and efficiently express proteins in eukaryotic cells, which is particularly important for protein replacement therapy.
04 Potential disease treatment applications
circRNA has shown potential in the treatment of various diseases, including rare diseases, cancer therapy, and as part of vaccine development.
05 Innovative delivery system
The newly developed delivery systems, such as lipid nanoparticles (LNPs), can effectively deliver circRNA to target cells, which is crucial for the targeting and efficacy of drugs.
06 Vaccine development
circRNA vaccines have shown potential in the development of COVID-19 vaccines, capable of inducing the production of more neutralizing antibodies and effective T-cell responses, and are more stable at ambient temperatures compared to linear mRNA, which helps simplify storage and transportation conditions.
07 Gene editing
circRNA also shows potential in the field of gene editing, capable of serving as a template for gene editing systems, providing more persistent and stable protein expression.
The circRNA production process includes: template preparation, in vitro transcription, removal of DNA template, circularization, and purification. After RNA circularization, un-circularized RNA needs to be digested with Ribonuclease R (RNase R) to enrich circRNA.
YEASEN has a bidirectional molecular enzyme rational design and directed evolution platform, as well as a molecular enzyme production base specifically designed for GMP-level products. Develop and produce GMP-grade RNase R products with high purity and strong digestive activity, providing support for the launch of new RNA vaccines/drugs.
Figure 1: Circular RNA production process diagram
(Source:Front Immunol 2023 Jan 12:13:1091797. doi: 10.3389/fimmu.2022.1091797. eCollection 2022.)
RNase R (Ribonuclease R, RNase R) is a Mg2+-dependent 3'→5' exoribonuclease originating from Escherichia coli. It can digest all linear RNA and has difficulty digesting circRNA, Lasso RNA, or double-stranded RNA molecules with less than 7 nucleotides protruding at the 3' end. RNase R is commonly used to remove linear RNA molecules, achieving the purpose of enriching non-linear RNAs such as circRNA and lariat RNA.
The main application regions are:
- Gene expression research;
- Research on alternative splicing;
- Enrich circRNA from biological samples;
- Identify intronic lariat structure RNA;
- Identify exon circRNA.
Product Features
- Protein purity (SDS-PAGE) ≥95%;
- No exonuclease residue;
- Compared to competitors, the product has equivalent capability in digesting linear RNA;
- Compared to competitors, the product has a similar effect on circRNA enrichment.
Performance demonstration
Protein purity (SDS-PAGE) ≥95%
Figure 2: Protein Purity Test: From left to right, the results are for different volumes (1 μL, 2 μL, 3 μL) of the same product concentration. The results show that the protein purity of YEASEN RNase R product is ≥95%.
No exonuclease residue
Figure 3: Residual Detection of Exonuclease: add 20 U of YEASEN RNase R to 0.5 μg of λDNA-Hind III digest, incubate at 37°C for 4 hours, and then perform agarose gel electrophoresis. The results show that the DNA bands did not change, indicating that there is no residual exonuclease in the YEASEN RNase R product.
Comparable to the A imported brand RNase R product in digesting linear RNA
Figure 4: Comparison of the ability of YEASEN RNase R products and A imported manufacturer's RNase R products to digest linear RNA:When the YEASEN RNase R product and the RNase R product from an imported manufacturer A were added to a reaction system containing linear RNA substrates, the results showed that when the amount of RNase R product added reached 2-4 U, the RNA bands became weak or even disappeared, indicating that RNase R has a digestive effect on linear RNA molecules. Moreover, the digestive capacity of YEASEN RNase R product and the RNase R product from the imported manufacturer A on linear RNA molecules is comparable.
Equivalent to the circRNA enrichment effect of A's imported brand RNase R product
Figure 5: Comparison of the enrichment effect of circRNA by YEASEN RNase R product and RNase R product from an imported manufacturer A: The reaction system containing circRNA substrates was subjected to reactions with YEASEN products and RNase R products from an imported manufacturer A, respectively. The results showed that there was no significant difference in the brightness of RNA bands after treatment with YEASEN's RNase R product and the RNase R product from the imported manufacturer A, indicating that circRNA can withstand digestion by RNase R. The above results confirm that both YEASEN products and A imported manufacturer's RNase R products can be effectively used for circRNA enrichment.
Customer feedback
YEASEN RNase R has better cleavage effects on linear RNA compared to other brands.
Figure 6: Comparison of the effects of RNase R digestion on linear mRNA molecules between YEASEN products and manufacturers A and B:The reaction was carried out by adding the YEASEN RNase R product and the RNase R products from manufacturers A and B to a reaction system containing 1 μg of linear mRNA substrate. The results showed that the YEASEN RNase R product had better digestion efficiency on linear RNA compared to the RNase R products from manufacturers A and B.
Product Information
|
Product Name |
Product Code |
Product Specifications |
|
RNase R GMP-grade (20 U/μL) |
14615ES |
500U/5000U/20KU/200KU |
|
10×RNase R Reaction Buffer GMP-grade |
14616ES |
500µl/1 mL/5 mL/10mL/100mL |
References
1.Qu L, Yi Z, Shen Y, et al. Circular RNA vaccines against SARS-CoV-2 and emerging variants. Cell. 2022;185(10):1728-1744.e16. doi:10.1016/j.cell.2022.03.044
2.Chen L, Wang C, Sun H, et al. The bioinformatics toolbox for circRNA discovery and analysis. Brief Bioinform. 2021;22(2):1706-1728. doi:10.1093/bib/bbaa001