Highly efficient reprogramming to pluripotency and directed differentiation of human cells using synthetic modified mRNA

By Luigi A. Warren, Philip D. Manos, Tim Ahfeldt, Yuin Han Loh, Hu Li, F. Lau, Wataru Ebina, P. Mandal, Zachary D. Smith, Alexander Meissner and 6 more. Cell Stem Cell, 2010

2,639 citations recorded by the source corpus.

SUMMARY Clinical application of induced pluripotent stem (iPS) cells is limited by the low efficiency of iPS derivation and the fact that most protocols modify the genome to effect cellular reprogramming. Moreover, safe and effective means of directing the fate of patient-specific iPS cells towards clinically useful cell types are lacking. Here we describe a simple, non-integrating strategy for reprogramming cell fate based on administration of synthetic mRNA modified to overcome innate anti-viral responses. We show that this approach can reprogram multiple human cell types to pluripotency with efficiencies that greatly surpass established protocols. We further show that the same technology can be used to efficiently direct the differentiation of RNA-induced pluripotent stem (RiPS) cells into terminally differentiated myogenic cells. This technology represents a safe, efficient strategy for somatic cell reprogramming and directing cell fate that has broad applicability for basic research, disease modeling and regenerative medicine.

Subfields: Molecular Biology

View Paper: https://www.semanticscholar.org/paper/c8119935f45fb4bf07f00a78cc78eb798e3a97c1

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In Molecular Biology

1. K. Hochedlinger
2. M. Stadtfeld
3. K. Karikó
4. Ugur Sahin
5. Ö. Türeci
6. D. Weissman
7. Chuan He
8. Hiromi Muramatsu

Citations in the corpus

• Gene therapy clinical trials worldwide to 2012 – an update (2013, background): “Originally produced by retroviral-mediated delivery, refinements to the system using non-integrating vectors and transient expression systems [77–81] will also address safety”
• Emerging trends and new developments in regenerative medicine: a scientometric update (2000 – 2014) (2014, neutral): “(2010) CELL STEM CELL, V7, P618 [45] 113.”
• mRNA-based therapeutics — developing a new class of drugs (2014, positive): “In 2010, it was demonstrated that IVT mRNA containing pseudouridine and 5-methylcytydine and encoding the Yamanaka stem cell factors 156 could be used as a safe strategy for efficiently reprogramming cells to pluripotency without leaving residual traces of transgenes 157 .”
• Induced pluripotency: history, mechanisms, and applications. (2010, background): “A more efficient and safer way of producing integration-free iPSCs may be the introduction of modified RNA molecules encoding for the reprogramming factors into somatic cells, which has been validated recently (Warren et al. 2010).”
• Widespread occurrence of 5-methylcytosine in human coding and non-coding RNA (2012, background): “More recently, it was reported that the methyl-CpG binding protein 2 (MECP2) associates with RNA and can regulate mRNA splicing (24,25) and that reprogramming of cells to pluripotency can be achieved using m(5)C and pseudouridine-modified mRNAs encoding the four Yamanaka factors (26).”
• Expression of therapeutic proteins after delivery of chemically modified mRNA in mice (2011, background): “Moreover, administration of chemically modified mRNA may have potential in the field of regenerative medicine including previously demonstrated reprogramming of stem cell”
• Highly efficient miRNA-mediated reprogramming of mouse and human somatic cells to pluripotency (2011, positive): “Thus, miR302/367 iPSC reprogramming is more efficient than previously described methods, including transfection of synthetic mRNAs for OSKM factors (Warren et al., 2010).”
• Reference Maps of Human ES and iPS Cell Variation Enable High-Throughput Characterization of Pluripotent Cell Lines (2011, positive): “For example, it will be interesting to measure whether the use of integration-free methods for reprogramming (Soldner et al., 2009; Warren et al., 2010) has an effect on the differentiation propensities of iPS cell lines.”