mRNA reprogramming technology offers a "footprint-free" method for producing induced pluripotent stem cells (iPSCs), ideal for clinical use. It is promising for disease modeling and regenerative medicine, with significant commercial potential. Creative Biolabs has developed a non-integrative RNA reprogramming platform using advanced technology to provide fast, safe, and efficient services, delivering high-quality iPSC cell derivation services tailored to specific needs.
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.
mRNA-based reprogramming has proven to be a non-viral, non-integrating method for reliable, safe and efficient generation of iPSC cell lines.
Fig.1 Principle workflow of mRNA-based reprogramming.1
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 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.
A: mRNA-based cell reprogramming is a non-integrating method to convert somatic cells into induced pluripotent stem cells (iPSCs) using mRNA to express reprogramming factors without altering the cellular DNA.
A: It is a footprint-free method that avoids genomic integration, reduces the risk of tumorigenicity, and eliminates the need for screening for residual vectors.
A: Creative Biolabs offers reprogramming services for T cells, dendritic cells, natural killer cells, B cells, and iPSCs.
A: It holds promise in disease modeling, regenerative medicine, gene therapy, and the generation of personalized stem cells.
A: Researchers can opt for tailored RNA reprogramming services with various differentiation strategies to meet specific application and program needs.
This article discusses the generation of induced pluripotent stem cells (iPSCs) using mRNA reprogramming methods. It focuses on creating cardiomyocytes from these iPSCs for cardiac regenerative therapy. The study outlines the process of generating footprint-free iPSCs and characterizing the resulting cardiomyocytes through molecular, structural, and functional analyses. The cardiomyocytes exhibited appropriate responses to pharmacological agents, indicating their potential for clinical applications in cardiac regenerative medicine. The study highlights the advantages of mRNA reprogramming, including high efficiency and absence of genomic integration, making it a promising approach for developing patient-specific iPSCs and cardiomyocytes.
Fig.2 Characterization of transgene free iPSC.2
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