Cited by 2,662Pluripotent Stem Cells ResearchCRISPR and Genetic EngineeringRNA Interference and Gene Delivery

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

Open access
Warren, Luigi, Manos, Philip D., Ahfeldt, Tim, Loh, Yuin-Han, Li, Hu, Lau, Frank, Ebina, Wataru, Mandal, Pankaj K. +8 more·2010·Cell Stem Cell

Abstract

Clinical application of induced pluripotent stem cells (iPSCs) is limited by the low efficiency of iPSC 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 iPSCs toward clinically useful cell types are lacking. Here we describe a simple, nonintegrating strategy for reprogramming cell fate based on administration of synthetic mRNA modified to overcome innate antiviral 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 cells (RiPSCs) 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.

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Crossref · 10.1016/j.stem.2010.08.012

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  • Nucleoside modifications can strongly enhance protein expression and suppress cytokine secretion.

    “Using enhanced green fluorescent protein mRNA, Rossi and colleagues confirmed that nucleoside modifications can strongly enhance protein expression and suppress cytokine secretion.”

    From Sequence-engineered mRNA without chemical nucleoside modifications enables an effective protein therapy in large animals (2015) · cites this paper as [24]

    In Results, “Development of modified-RNAs for directing cell fate” (page unavailable), the paper reports that nucleoside modifications in GFP-encoding mRNA markedly increased protein expression and dramatically attenuated interferon signaling. It assessed interferon-response gene expression by qRT-PCR, not cytokine secretion, so the secretion claim is not established by our copy. (LLM, conf=0.87)

    Experimental — accuracy not yet measured
  • Conventional unmodified mRNAs are inferior to molecules harboring specific nucleoside modifications, but the picture changes dramatically with sophisticated sequence-engineering.

    “Our work principally confirms that conventional unmodified mRNAs are inferior to molecules harboring specific nucleoside modifications as suggested by various recent reports,23,24,25,26,27 however, the picture changes dramatically with a sophisticated sequence-engineering.”

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Authors
Warren, L., Manos, P. D., Ahfeldt, T., Loh, Y.-H., Li, H., Lau, F., Ebina, W., Mandal, P. K., Smith, Z. D., Meissner, A., Daley, G. Q., Brack, A. S., Collins, J. J., Cowan, C., Schlaeger, T. M., & Rossi, D. J.
Affiliations
  • Boston Children's Hospital (US)
  • Harvard University (US)
  • Boston University (US)
  • Broad Institute (US)
  • Brigham and Women's Hospital (US)
Corresponding author
Rossi, Derrick J.
Venue
Cell Stem Cell
Publisher
Elsevier BV
Published
2010-11
Volume
7
Issue
5
Pages
618-630
ISSN
  • 1875-9777 (ISSN-L)
  • 1934-5909
OpenAlex
W2098597036
Cited by
2,662
References
55
Citations by year
2026: 74 · 2025: 67 · 2024: 89 · 2023: 81 · 2022: 109 · 2021: 153 · 2020: 148 · 2019: 130 · 2018: 178 · 2017: 167 · 2016: 201 · 2015: 208 · 2014: 235 · 2013: 264 · 2012: 288
FWCI
106.76
Type
journal article
Language
en
Open access
Locations
  • Cell stem cell (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
  • Digital Access to Scholarship at Harvard (DASH) (Harvard University) (submittedVersion, cc-by)
Funders
  • Howard Hughes Medical Institute
  • National Heart, Lung, and Blood Institute (U01HL100001)
  • National Institute of Dental and Craniofacial Research (RL1DE01902, RL1DE019021)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
Keywords
synthetic mRNA, somatic cell reprogramming, pluripotency, directed differentiation, myogenesis, antiviral response, disease modeling, regenerative medicine
MeSH terms
Cell Differentiation, Cells, Cultured, Humans, RNA, Messenger, Cell Lineage, Induced Pluripotent Stem Cells, Cellular Reprogramming

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