Custom mRNA-based Cell Reprogramming Service

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.

Introduction of mRNA Reprogramming

Traditional generation of iPSCs has relied on the expression of inducible embryonic gene expression profiling factors by integrating viral gene expression vectors. This could lead to potentially oncogenic alterations of the normal genome. mRNA reprogramming is a “non-integrating” reprogramming system that has shown several significant advantages over other reprogramming methods.

• mRNA reprogramming is a "footprint-free" method that will not result in heritable changes to cellular DNA and an unacceptable risk of tumorigenicity. When transfected mRNA is used to express reprogramming factors (such as OCT4, KLF4, SOX2, and cMYC), there's no need to wait a long time for vectors to clear, no need to screen iPSCs for sporadic genomic integration or other vector traces, and no residual risk associated with the possibility of false negatives during the screening process.
• By employing self-replicating vectors that give a prolonged burst of transgene expression after being introduced into target cells in a single delivery step, mRNA reprogramming offers the convenience of "single-shot" reprogramming with long-lasting transgene expression.

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.

• Custom T Cell Reprogramming by mRNA

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.

• Custom Dendritic Cell Reprogramming by mRNA

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).

• Custom Natural Killer Cell Reprogramming by mRNA

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.

• Custom B Cell Reprogramming by mRNA

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.

• Custom iPS Cell Reprogramming by mRNA

Reprogramming with synthetic capped mRNAs holds great promise for the safe generation of human-derived iPSCs. With the advantages of high efficiency (up to 4.4%), low activation against an innate antiviral response, and generation of high-quality clinically relevant iPS, it has been the most promising approach.

Highlighted Features

• Custom-tailored RNA reprogramming services (integration-free, non-viral, safe and efficient)
• Optional differentiation strategies (readily available cell resources with flexible, efficient and iterative custom solutions to meet specific application and program needs)
• Rigorous quality control and potent technical support

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.

Reference

1. Barahona, D. E.; et al. A review of cellular reprogramming: limitations and recent advances. Bionatura. 2018, 3(2).