mRNA reprogramming technology is the most unambiguously "footprint-free", most productive, and most suitable method for clinical production of induced pluripotent stem cells (iPSCs). It holds great promise in the areas of disease modeling and regenerative medicine and shows unique commercial potential. With access to advanced instruments, fit-for-purpose laboratories, and enriched expertise, Creative Biolabs has developed a non-integrative RNA reprogramming platform to offer a fast, safe and efficient RNA reprogramming system to deliver high-quality iPSCs cell lines to meet your specific needs.
Conventional generation of iPSCs is dependent on integrating viral gene expression vectors to express the factors that induce an embryonic gene expression profile, which may cause potentially oncogenic alteration of the normal genome. mRNA reprogramming is a “non-integrating” reprogramming system that has shown several significant advantages over other reprogramming methods.
Fig.1 Principle workflow of mRNA-based reprogramming. (Barahona, 2018)
mRNA-based reprogramming has proven to be a non-viral, non-integrating method for reliable, safe and efficient generation of iPSC cell lines.
It has been demonstrated that T cells are amenable to reprogramming by overexpressing Oct4, Sox2, Klf4, and Myc with the ectopic expression of p53 knockdown. mRNA-based reprogramming method contributes to the generation of iPSCs derived from T cells and advances the improvement of efficient T cell reprogramming and gene therapy approaches.
Synthetic mRNA has emerged as a powerful tool for the transfer of reprogramming factors into dendritic cells (DCs) in order to induce pluripotency. The established iPSC-promoting transcription factors include OCT4, SOX2, KLF4, cMYC, NANOG, and LIN28 (OSKMNL).
Natural killer (NK) cells can be genetically reprogramed efficiently using a synthetic mRNA electroporation method that induces rapid and reproducible transgene expression (such as chemokine receptor CCR7, high-affinity antibody-binding receptor CD16, etc.) with high efficiency, without negatively influencing their viability, phenotype, and cytotoxic function.
B cells can be reprogramed by mRNA through the transfection of synthetic mRNA encoding different transcription factors. For early-stage non-terminally differentiated B cells, Yamanaka factors OSKM (OCT4, SOX2, KLF4, cMYC) can be used to dedifferentiate B cells into a pluripotent state. For mature late-stage B cells, it requires supplementary transcriptional factors, such as CCAAT/enhancer-binding-protein-alpha (a myeloid transcription factor) or specific knockdown of the B cell transcription factor PAX5.
Reprogramming with synthetic capped mRNAs holds great promise for the safe generation of iPSCs of human origin. It has been the most promising approach due to its relatively high efficiency (up to 4.4%), low activation of an innate antiviral response, and the production of high-quality, clinically relevant iPSs.
Creative Biolabs is guaranteed to provide highly efficient iPSC cell derivation services of different cells with our advanced technology, including reprogramming factor delivery by mRNA. If you are interested in our services, please feel free to contact us.