The mission of CELLSCRIPT™ is to provide the best products and technologies for making and using RNA for translation in cells for clinical research and therapeutics.
Current products include kits for in vitro transcription, 5' RNA capping using either a cap analog or capping enzymes, and 3' RNA polyadenylation, as well as all-in-one kits for making capped, poly(A)-tailed mRNA for translation in cells.
CELLSCRIPT™ recently introduced INCOGNITO™ RNA Transcription Kits for in vitro synthesis of RNA that contains modified nucleosides, such as pseudouridine (Ψ) and/or 5-methylcytidine (m5C) in place of the corresponding U or C canonical nucleosides (Figure 1). These kits are so-named because the capped, polyadenylated and nucleoside-modified RNA products (called "INCOGNITO mRNAs") are disguised so they do not induce innate immune responses to the same extent as the corresponding unmodified mRNAs when transfected into mammalian cells that express a variety of RNA sensors.
As documented in journal articles1–3 and patent applications,4 the technologies on which INCOGNITO RNA kits and other products are based were invented by Professors Katalin Karikó and Drew Weissman at the University of Pennsylvania and exclusively licensed to CELLSCRIPT™ for all fields of use. Drs. Weissman and Karikó showed that INCOGNITO mRNA, besides being less immunogenic, is translated into protein at much higher levels than the corresponding mRNA that does not contain modified nucleosides, both in cultured cells (Figure 2) and in whole organisms (Figure 3). INCOGNITO-type Ψ- and m5C-modified mRNA encoding KLF4, LIN28, cMYC, OCT4 and SOX2 that was repeatedly transfected into somatic cells, such as fibroblasts and keratinocytes, resulted in highly efficient generation of induced pluripotent stem cells.5
The delivery of protein-encoding INCOGNITO mRNA to cells in culture or in vivo to an organism has the potential to produce a therapeutic effect by compensating for a missing or defective protein, overexpressing a desired protein, inducing a change in cellular phenotype, or triggering a disease-specific immune response. Thus, we believe INCOGNITO mRNAs will have many uses in regenerative medicine, such as for: cell reprogramming; cell therapies; cell, tissue or organ transplantation or repair; tissue or organ engineering; enzyme replacement therapies; and immunotherapies or immunomodulation therapies.
Contact CELLSCRIPT™ to discuss kits for making research or therapeutic grades of INCOGNITO mRNA or unmodified RNA, or possible custom manufacturing of an appropriate grade and scale of RNA for your project.
- Karikó K, Buckstein M, Ni H, Weissman D. Immunity 23: 165–75, 2005. Suppression of RNA recognition by Toll-like receptors: The impact of nucleoside modification and the evolutionary origin of RNA. Journal Article
- Karikó K, Muramatsu H, Welsh FA, Ludwig J, Kato H, Akira S, Weissman D. Molecular Therapy 16: 1833–40, 2008. Incorporation of pseudouridine into mRNA yields superior nonimmunogenic vector with increased translational capacity and biological stability. Journal Article
- Anderson B. Muramatsu H, Nallagatla SR, Bevilacqua P. Sansing L, Weissman D, Karikó K. Nucleic Acids Res 38: 5884–92, 2010. Incorporation of pseudouridine into mRNA enhances translation by diminishing PKR activation. Journal Article
- E.g., United States Patent Application No. US 20090286852 Journal Article
- Warren L, Manos PD, Ahfeldt T, Loh YH, Li H, Lau F, Ebina W,
Mandal PK, Smith ZD, Meissner A, Daley GQ, Brack AS, Collins JJ,
Cowan C, Schlaeger TM, Rossi DJ. Cell Stem Cell.
7(5): 549–50, 2010.