Mitigating off-target effects in CRISPR/Cas9-mediated in vivo gene editing

J Mol Med (Berl). 2020 May;98(5):615-632. doi: 10.1007/s00109-020-01893-z. Epub 2020 Mar 20.


The rapid advancement of genome editing technologies has opened up new possibilities in the field of medicine. Nuclease-based techniques such as the CRISPR/Cas9 system are now used to target genetically linked disorders that were previously hard-to-treat. The CRISPR/Cas9 gene editing approach wields several advantages over its contemporary editing systems, notably in the ease of component design, implementation and the option of multiplex genome editing. While results from the early phase clinical trials have been encouraging, the small patient population recruited into these trials hinders a conclusive assessment on the safety aspects of the CRISPR/Cas9 therapy. Potential safety concerns include the lack of fidelity in the CRISPR/Cas9 system which may lead to unintended DNA modifications at non-targeted gene loci. This review focuses modifications to the CRISPR/Cas9 components that can mitigate off-target effects in in vitro and preclinical models and its translatability to gene therapy in patient populations.

Keywords: CRISPR/Cas9; Clinical trials; Gene therapy; In vivo; Off-target effects.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Animals
  • CRISPR-Cas Systems*
  • DNA Repair
  • Disease Management
  • Gene Editing*
  • Gene Expression Regulation
  • Gene Targeting
  • Genetic Predisposition to Disease
  • Genetic Therapy* / methods
  • Humans
  • MicroRNAs / genetics
  • Models, Animal
  • Organ Specificity / genetics
  • Promoter Regions, Genetic
  • RNA, Guide


  • MicroRNAs
  • RNA, Guide