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  3. Small molecules enhance CRISPR genome editing in pluripotent stem cells

Small molecules enhance CRISPR genome editing in pluripotent stem cells

  • Cell Stem Cell. 2015 Feb 5;16(2):142-7. doi: 10.1016/j.stem.2015.01.003.
Chen Yu 1,  Yanxia Liu 2,  Tianhua Ma 1,  Kai Liu 1,  Shaohua Xu 1,  Yu Zhang 1,  Honglei Liu 3,  Marie La Russa 4,  Min Xie 1,  Sheng Ding 5,  Lei S Qi 6
Affiliations

Affiliations

  • 1 The Gladstone Institute of Cardiovascular Disease, 1650 Owens Street, San Francisco, CA 94158, USA.
  • 2 Department of Bioengineering, Stanford University, Stanford, CA 94305, USA.
  • 3 Department of Bioengineering, Stanford University, Stanford, CA 94305, USA; Bioinformatics Division, Center for Synthetic and Systems Biology, TNLIST/Department of Automation, Tsinghua University, Beijing 100084, China.
  • 4 Department of Bioengineering, Stanford University, Stanford, CA 94305, USA; Biomedical Sciences Graduate Program, University of California, San Francisco, 600 16(th) Street, San Francisco, CA 94158, USA.
  • 5 The Gladstone Institute of Cardiovascular Disease, 1650 Owens Street, San Francisco, CA 94158, USA; Department of Pharmaceutical Chemistry, University of California, San Francisco, 600 16(th) Street, San Francisco, CA 94158, USA. Electronic address: sheng.ding@gladstone.ucsf.edu.
  • 6 Department of Bioengineering, Stanford University, Stanford, CA 94305, USA; Department of Chemical and Systems Biology, Stanford University, Stanford, CA 94305, USA; ChEM-H, Stanford University, Stanford, CA 94305, USA. Electronic address: stanley.qi@stanford.edu.
PMID: 25658371 DOI: 10.1016/j.stem.2015.01.003
Abstract

The Bacterial CRISPR-Cas9 system has emerged as an effective tool for sequence-specific gene knockout through non-homologous end joining (NHEJ), but it remains inefficient for precise editing of genome sequences. Here we develop a reporter-based screening approach for high-throughput identification of chemical compounds that can modulate precise genome editing through homology-directed repair (HDR). Using our screening method, we have identified small molecules that can enhance CRISPR-mediated HDR efficiency, 3-fold for large fragment insertions and 9-fold for point mutations. Interestingly, we have also observed that a small molecule that inhibits HDR can enhance frame shift insertion and deletion (indel) mutations mediated by NHEJ. The identified small molecules function robustly in diverse cell types with minimal toxicity. The use of small molecules provides a simple and effective strategy to enhance precise genome engineering applications and facilitates the study of DNA repair mechanisms in mammalian cells.

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