title: Small molecules enhance CRISPR genome editing in pluripotent stem cells.
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.
authors: The Gladstone Institute of Cardiovascular Disease, 1650 Owens Street, San Francisco, CA 94158, USA.
authors: Chen
authors: C
authors: Yu
authors: Department of Bioengineering, Stanford University, Stanford, CA 94305, USA.
authors: Yanxia
authors: Y
authors: Liu
authors: The Gladstone Institute of Cardiovascular Disease, 1650 Owens Street, San Francisco, CA 94158, USA.
authors: Tianhua
authors: T
authors: Ma
authors: The Gladstone Institute of Cardiovascular Disease, 1650 Owens Street, San Francisco, CA 94158, USA.
authors: Kai
authors: K
authors: Liu
authors: The Gladstone Institute of Cardiovascular Disease, 1650 Owens Street, San Francisco, CA 94158, USA.
authors: Shaohua
authors: S
authors: Xu
authors: The Gladstone Institute of Cardiovascular Disease, 1650 Owens Street, San Francisco, CA 94158, USA.
authors: Yu
authors: Y
authors: Zhang
authors: 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.
authors: Honglei
authors: H
authors: Liu
authors: 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.
authors: Marie
authors: M
authors: La Russa
authors: The Gladstone Institute of Cardiovascular Disease, 1650 Owens Street, San Francisco, CA 94158, USA.
authors: Min
authors: M
authors: Xie
authors: 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.
authors: Sheng
authors: S
authors: Ding
authors: 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.
authors: Lei S
authors: LS
authors: Qi
mesh_terms: D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D004260:DNA Repair / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D005818:Genetic Engineering / Q000379:methods*; D015894:Genome, Human / Q000235:genetics*; D006367:HeLa Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D015394:Molecular Structure; D039904:Pluripotent Stem Cells / Q000187:drug effects* / Q000378:metabolism*; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology*; D013329:Structure-Activity Relationship
chemical_list: D054852:Small Molecule Libraries