mRNA-Based iPSC Production
The discovery of induced pluripotent stem cells (iPSCs) was a remarkable achievement in biotechnology. The derivation of iPSCs is paramount for the development of disease modeling, enhanced platforms for drug screening, and widespread use of cell transplantation therapy in regenerative medicine. mRNA reprogramming technology serves as a fast, safe, and efficient method that generates clinical-grade iPSCs from somatic tissue.
The generation of iPSCs
In 2006, the reprogramming of somatic cells to iPSCs using a specific set of transcription factors (Yamanaka factors, including Oct4, Sox2, Klf4, and cMyc) was first described by Yamanaka. The discovery of iPSCs is groundbreaking in biotechnology. Because these iPSCs are a promising alternative to embryonic stem cells (ESCs) due to their products without destroying an embryo and this donor-specific isolation of somatic cells is considered safe in ethics. Since the discovery that human iPSCs can be produced by inducing the expression of Yamanaka factors, many different reprogramming techniques for generating iPSCs have emerged. Currently, these methods include DNA virus systems (retroviral and lentiviral vector), RNA virus system (Sendai virus), the use of episomal DNA plasmids, direct protein delivery of Yamanaka factors, and the synthetic mRNA transfection. Retroviral vectors used to deliver reprogramming factors into the cells are initial and commonly used methods. However, this method would lead to random integration of reprogramming factors into the genome. The use of episomal DNA plasmids and Sendai virus belongs to non-integrative methods and can be used for a variety of cell types. But they require monitoring for vector or viral clearance. As for direct protein delivery of Yamanaka factors, the method shows no risk of genome insertion. But it is limited for protein-based reprogramming protocols from most applications due to its low efficiency and a longer workflow. In particular, synthetic mRNA transfection is a promising and powerful reprogramming tool to convert somatic cells to iPSCs. Synthetic mRNA has a number of advantages over traditional approaches, including efficient and rapid-acting, integration-free, footprint-free, and allowing fine titration of gene dosage.
iPSC generation using mRNA
At present, mRNA-based reprogramming technology as a non-viral, non-integrating method, has proven to generate the reliable, safe, and efficient generation of transgene-free iPSCs from somatic tissue. It is reported that the reprogramming efficiency of this technology is greater than 1%, which is more efficient than other transfection methods.
Benefits of using the mRNA reprogramming technology compared to other reprogramming methods.
|Reprogramming protocols||DNA virus
|The use of episomal DNA plasmids||Recombinant proteins||mRNA-based reprogramming technology|
|Time course for colony isolation (weeks)||2-4||4||-||8||2|
|Risk of genome integration and mutagenesis||Yes||No||No||No||No|
Custom iPSC reprogramming by mRNA
Creative Biogene provides rapid, low-cost, and high-quality iPSCs from somatic cells based on an advanced form of mRNA reprogramming. Based on our advanced platform and expertise, our specialists are committed to providing an accurate and reliable generation of human iPSCs for drug discovery and other pre-clinical applications. We have put much effort into the optimization of mRNA-based reprogramming protocols in terms of various parameters, such as the numbers of transfections, different modifications of mRNAs, multiple vectors, and routes, as well as cell culture conditions. And finally, we have established a relatively simple process and an optimal protocol.
Fig1. Custom mRNA-based reprogramming
Features of our services
- Simple, safe, non-integrating mRNA reprogramming technology.
- Higher efficiencies than traditional approaches.
- Reprogram multiple human cell types, including skin fibroblasts, blood cells, and epithelial cells.
- Rigorous quality control protocols
Your success is important to us, and our custom iPSC generation service is supported by years of experience in mRNA-based reprogramming. Simply let us know about your research needs. We will propose the best strategy to best match your research objectives. Please feel free to contact us for more information.
- Badieyan, Z. S., & Evans, T. (2019). "Concise review: application of chemically modified mRNA in cell fate conversion and tissue engineering." Stem cells translational medicine, 8(8), 833-843.