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Stem Cells and Aging pp 337–346Cite as

CRISPR Base Editing in Induced Pluripotent Stem Cells

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2045))

Abstract

Induced pluripotent stem cells (iPSCs) have demonstrated tremendous potential in numerous disease modeling and regenerative medicine-based therapies. The development of innovative gene transduction and editing technologies has further augmented the potential of iPSCs. Cas9-cytidine deaminases, for example, have developed as an alternative strategy to integrate single-base mutations (C → T or G → A transitions) at specific genomic loci. In this chapter, we specifically describe CRISPR (clustered regularly interspaced short palindromic repeats) base editing in iPSCs for editing precise locations in the genome. This state-of-the-art approach enables highly efficient and accurate modifications in genes. Thus, this technique not only has the potential to have biotechnology and therapeutic applications but also the ability to reveal underlying mechanisms regarding pathologies caused by specific mutations.

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Acknowledgments

The Jonas Children’s Vision Care and Bernard & Shirlee Brown Glaucoma Laboratory are supported by the National Institutes of Health [P30EY019007, R01EY018213, §R01EY024698, R01EY026682, R21AG050437], National Cancer Institute Core [5P30CA013696], Foundation Fighting Blindness [TA-NMT-0116-0692-COLU], the Research to Prevent Blindness (RPB) Physician-Scientist Award, and unrestricted funds from RPB, New York, NY, USA. S.H.T. is a member of the RD-CURE Consortium and is supported by Kobi and Nancy Karp, the Crowley Family Fund, the Rosenbaum Family Foundation, the Tistou and Charlotte Kerstan Foundation, the Schneeweiss Stem Cell Fund, New York State [C029572], and the Gebroe Family Foundation. YJC and CLX contributed equally to this work. YJC and CLX wrote and edited the manuscript. XC and YTT were responsible for developing and finalizing the protocol. AGB, VBM, and SHT oversaw the writing process.

Conflict of Interest: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Author information

Author notes

  1. Ya-Ju Chang and Christine L. Xu contributed equally to this work and are co-first authors.

Authors and Affiliations

  1. Department of Ophthalmology, Columbia University, New York, NY, USA

    Ya-Ju Chang, Christine L. Xu, Xuan Cui, Yi-Ting Tsai & Stephen H. Tsang

  2. Jonas Children’s Vision Care, Bernard and Shirlee Brown Glaucoma Laboratory, Columbia University, New York, NY, USA

    Ya-Ju Chang, Christine L. Xu, Xuan Cui & Stephen H. Tsang

  3. Department of Pediatrics, University of Iowa, Iowa City, IA, USA

    Alexander G. Bassuk

  4. Department of Neurology, University of Iowa, Iowa City, IA, USA

    Alexander G. Bassuk

  5. Palo Alto Veterans Administration, Palo Alto, CA, USA

    Vinit B. Mahajan

  6. Omics Lab, Department of Ophthalmology, Byers Eye Institute, Stanford University, Palo Alto, CA, USA

    Vinit B. Mahajan

  7. Institute of Human Nutrition, College of Physicians and Surgeons, Columbia University, New York, NY, USA

    Yi-Ting Tsai & Stephen H. Tsang

  8. Department of Pathology and Cell Biology, Columbia University, New York, NY, USA

    Stephen H. Tsang

Authors
  1. Ya-Ju Chang
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  2. Christine L. Xu
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  3. Xuan Cui
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  4. Alexander G. Bassuk
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  5. Vinit B. Mahajan
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  6. Yi-Ting Tsai
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  7. Stephen H. Tsang
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Editor information

Editors and Affiliations

  1. Ottawa, ON, Canada

    Kursad Turksen

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© 2019 Springer Science+Business Media New York

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Cite this protocol

Chang, YJ. et al. (2019). CRISPR Base Editing in Induced Pluripotent Stem Cells. In: Turksen, K. (eds) Stem Cells and Aging . Methods in Molecular Biology, vol 2045. Humana, New York, NY. https://doi.org/10.1007/7651_2019_243

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  • DOI: https://doi.org/10.1007/7651_2019_243

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9712-1

  • Online ISBN: 978-1-4939-9713-8

  • eBook Packages: Springer Protocols

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