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CRISPR Base Editing in Induced Pluripotent Stem Cells

  • Ya-Ju Chang
  • Christine L. Xu
  • Xuan Cui
  • Alexander G. Bassuk
  • Vinit B. Mahajan
  • Yi-Ting Tsai
  • Stephen H. Tsang
Protocol
Part of the Methods in Molecular Biology book series (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.

Keywords

Base editing iPS cells Target-AID Cas9 Precise gene editing 

Notes

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.

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Copyright information

© Springer Science+Business Media New York 2019

Authors and Affiliations

  • Ya-Ju Chang
    • 1
    • 2
  • Christine L. Xu
    • 1
    • 2
  • Xuan Cui
    • 1
    • 2
  • Alexander G. Bassuk
    • 3
    • 4
  • Vinit B. Mahajan
    • 5
    • 6
  • Yi-Ting Tsai
    • 1
    • 7
  • Stephen H. Tsang
    • 1
    • 2
    • 7
    • 8
  1. 1.Department of OphthalmologyColumbia UniversityNew YorkUSA
  2. 2.Jonas Children’s Vision Care, Bernard and Shirlee Brown Glaucoma LaboratoryColumbia UniversityNew YorkUSA
  3. 3.Department of PediatricsUniversity of IowaIowa CityUSA
  4. 4.Department of NeurologyUniversity of IowaIowa CityUSA
  5. 5.Palo Alto Veterans AdministrationPalo AltoUSA
  6. 6.Omics Lab, Department of Ophthalmology, Byers Eye InstituteStanford UniversityPalo AltoUSA
  7. 7.Institute of Human Nutrition, College of Physicians and SurgeonsColumbia UniversityNew YorkUSA
  8. 8.Department of Pathology and Cell BiologyColumbia UniversityNew YorkUSA

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