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Effects of clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) protein 9 system-Based Deletion of miR-451 in Mouse Embryonic Stem Cells on Their Self-Renewal and Hematopoietic Differentiation

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Tissue Engineering and Regenerative Medicine Aims and scope

Abstract

Pluripotent stem cells (PSCs) are a useful source of cells for exploring the role of genes related with early developmental processes and specific diseases due to their ability to differentiate into all somatic cell types. Recently, the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) protein 9 system has proven to be a robust tool for targeted genetic modification. Here, we generated miR-451-deficient PSCs using the CRISPR/Cas9 system with PCR-based homologous recombination donor and investigated the impact of its deletion on self-renewal and hematopoietic development. CRISPR/Cas9-mediated miR-451 knockout did not alter the gene expressions of pluripotency, cellular morphology, and cell cycle, but led to impaired erythrocyte development. These findings propose that a combination of PSCs and CRISPR/Cas9 system could be useful to promote biomedical applications of PSCs by elucidating the function and manipulating of specific miRNAs during lineage specification and commitment.

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Acknowledgements

This study was supported by a grant from the Medical Research Center (2008-0062287) and the Basic Research Lab Program (2015R1A4A1038666) funded by the NRF of the Ministry of Science, ICT & Future Planning, Republic of Korea.

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    Authors and Affiliations

    1. Department of Biomedical Science, Graduate School of Biomedical Science and Engineering, Hanyang University, 222 Wangsimni-ro, Seongdong-gu, Seoul, 04763, South Korea

      Su-Jin Kim, Chang-Hoon Kim & Kye-Seong Kim

    2. College of Medicine, Hanyang University, Seoul, South Korea

      Kye-Seong Kim

    3. Department of Internal Medicine, School of Medicine, Kangwon National University, Kangwondaehakgil-1, Chuncheon, Gangwon-do, 24341, South Korea

      Borim An & Seok-Ho Hong

    4. Department of Molecular and Cellular Biochemistry, School of Medicine, Kangwon National University, Chuncheon, 24341, South Korea

      Kwon-Soo Ha

    5. Stem Cell Institute, Kangwon National University, Chuncheon, 24341, South Korea

      Seok-Ho Hong

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    1. Su-Jin Kim
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    Correspondence to Seok-Ho Hong or Kye-Seong Kim.

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    There are no human or animal experiments carried out for this article.

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    Su-Jin Kim and Chang-Hoon Kim have contributed equally to this work.

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    Kim, SJ., Kim, CH., An, B. et al. Effects of clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) protein 9 system-Based Deletion of miR-451 in Mouse Embryonic Stem Cells on Their Self-Renewal and Hematopoietic Differentiation. Tissue Eng Regen Med 14, 179–185 (2017). https://doi.org/10.1007/s13770-017-0031-8

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