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Methods Mol Biol. 2019 Jun 28. doi: 10.1007/7651_2019_243. [Epub ahead of print]

CRISPR Base Editing in Induced Pluripotent Stem Cells.

Chang YJ1,2, Xu CL1,2, Cui X1,2, Bassuk AG3,4, Mahajan VB5,6, Tsai YT1,7, Tsang SH8,9,10,11.

Author information

1
Department of Ophthalmology, Columbia University, New York, NY, USA.
2
Jonas Children's Vision Care, Bernard and Shirlee Brown Glaucoma Laboratory, Columbia University, New York, NY, USA.
3
Department of Pediatrics, University of Iowa, Iowa City, IA, USA.
4
Department of Neurology, University of Iowa, Iowa City, IA, USA.
5
Palo Alto Veterans Administration, Palo Alto, CA, USA.
6
Omics Lab, Department of Ophthalmology, Byers Eye Institute, Stanford University, Palo Alto, CA, USA.
7
Institute of Human Nutrition, College of Physicians and Surgeons, Columbia University, New York, NY, USA.
8
Department of Ophthalmology, Columbia University, New York, NY, USA. crispr.ips@gmail.com.
9
Jonas Children's Vision Care, Bernard and Shirlee Brown Glaucoma Laboratory, Columbia University, New York, NY, USA. crispr.ips@gmail.com.
10
Institute of Human Nutrition, College of Physicians and Surgeons, Columbia University, New York, NY, USA. crispr.ips@gmail.com.
11
Department of Pathology and Cell Biology, Columbia University, New York, NY, USA. crispr.ips@gmail.com.

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; Cas9; Precise gene editing; Target-AID; iPS cells

PMID:
31250381
DOI:
10.1007/7651_2019_243

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