Optimization of Synthetic mRNA for Highly Efficient Translation and its Application in the Generation of Endothelial and Hematopoietic Cells from Human and Primate Pluripotent Stem Cells

Kran Suknuntha, Lihong Tao, Vera Brok-Volchanskaya, Saritha S D'Souza, Akhilesh Kumar, Igor Slukvin

Research output: Contribution to journalArticlepeer-review

23 Scopus citations

Abstract

Identification of transcription factors that directly convert pluripotent stem cells (PSCs) into endothelial and blood cells and advances in the chemical modifications of messenger RNA (mRNA) offer alternative nucleic acid-based transgene-free approach for scalable production of these cells for drug screening and therapeutic purposes. Here we evaluated the effect of 5' and 3' RNA untranslated regions (UTRs) on translational efficiency of chemically-modified synthetic mRNA (modRNA) in human PSCs and showed that an addition of 5'UTR indeed enhanced protein expression. With the optimized modRNAs expressing ETV2 or ETV2 and GATA2, we are able to produce VE-cadherin+ endothelial cells and CD34+CD43+ hematopoietic progenitors, respectively, from human PSCs as well as non-human primate (NHP) PSCs. Overall, our findings provide valuable information on the design of in vitro transcription templates being used in PSCs and its broad applicability for basic research, disease modeling, and regenerative medicine.

Original languageEnglish (US)
Pages (from-to)525-534
Number of pages10
JournalStem Cell Reviews
Volume14
Issue number4
DOIs
StatePublished - Aug 2018
Externally publishedYes

Bibliographical note

Funding Information:
Acknowledgements We thank the Cell Processing and Manipulation Core in the Translational Cores, and Physicians and Nurses at University of Wisconsin Carbone Cancer Center and Cincinnati Children’s Hospital Medical Center for obtaining and processing bone marrow samples and Translational Research Trials Office for providing the regulatory and administrative support for this endeavor. This work was supported by funds from the National Institute of Health (1R01HL132891, 4R01HL116221, and P51OD011106).

Funding Information:
We thank the Cell Processing and Manipulation Core in the Translational Cores, and Physicians and Nurses at University of Wisconsin Carbone Cancer Center and Cincinnati Children?s Hospital Medical Center for obtaining and processing bone marrow samples and Translational Research Trials Office for providing the regulatory and administrative support for this endeavor. This work was supported by funds from the National Institute of Health (1R01HL132891, 4R01HL116221, and?P51OD011106).

Publisher Copyright:
© 2018, Springer Science+Business Media, LLC, part of Springer Nature.

Keywords

  • 5' Untranslated Regions/genetics
  • Animals
  • Antigens, CD/genetics
  • Antigens, CD34/genetics
  • Cadherins/genetics
  • Cell Differentiation/genetics
  • Cell Line
  • Cells, Cultured
  • Endothelial Cells/cytology
  • GATA2 Transcription Factor/genetics
  • Hematopoietic Stem Cells/cytology
  • Humans
  • Leukosialin/genetics
  • Pluripotent Stem Cells/cytology
  • Primates
  • Protein Biosynthesis
  • RNA, Messenger/genetics
  • Transcription Factors/genetics

PubMed: MeSH publication types

  • Journal Article
  • Research Support, N.I.H., Extramural

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