CRISPR Base Editing in Induced Pluripotent Stem Cells

Methods Mol Biol. 2019:2045:337-346. doi: 10.1007/7651_2019_243.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • CRISPR-Cas Systems / genetics*
  • Cell Cycle
  • Cells, Cultured
  • Cytidine Deaminase / genetics
  • Cytidine Deaminase / metabolism*
  • Cytosine / chemistry
  • Cytosine / metabolism
  • Electroporation
  • Gene Editing / instrumentation
  • Gene Editing / methods*
  • Genome, Human / drug effects
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Induced Pluripotent Stem Cells / chemistry
  • Induced Pluripotent Stem Cells / cytology
  • Induced Pluripotent Stem Cells / drug effects
  • Induced Pluripotent Stem Cells / metabolism*
  • Software
  • Transfection / methods
  • Uracil / chemistry

Substances

  • Uracil
  • Cytosine
  • Cytidine Deaminase