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

doi:10.1007/s12015-018-9805-1

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 journal › Article › peer-review

19 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 language	English (US)
Pages (from-to)	525-534
Number of pages	10
Journal	Stem Cell Reviews
Volume	14
Issue number	4
DOIs	https://doi.org/10.1007/s12015-018-9805-1
State	Published - Aug 2018
Externally published	Yes

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

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1007/s12015-018-9805-1

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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. / Suknuntha, Kran; Tao, Lihong; Brok-Volchanskaya, Vera et al.

In: Stem Cell Reviews, Vol. 14, No. 4, 08.2018, p. 525-534.

Research output: Contribution to journal › Article › peer-review

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title = "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",

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.",

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author = "Kran Suknuntha and Lihong Tao and Vera Brok-Volchanskaya and D'Souza, {Saritha S} and Akhilesh Kumar and Igor Slukvin",

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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

Abstract

Bibliographical note

Keywords

PubMed: MeSH publication types

UN SDGs

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