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Publications

Journal Article Review Epublish

PubMed: 40777742 DOI: 10.1016/j.omtn.2025.102636

Off-target effects in CRISPR-Cas genome editing for human therapeutics: Progress and challenges.

Mol Ther Nucleic Acids 36(3):102636

Authors: Nechama Kalter, Carla Fuster-García, Alfredo Silva, Víctor Ronco-Díaz, Stefano Roncelli, Giandomenico Turchiano, Jan Gorodkin, Toni Cathomen, Karim Benabdellah, Ciaran Lee, Ayal Hendel

Affiliations

  • The Institute for Advanced Materials and Nanotechnology, The Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan 5290002, Israel.
  • Institute for Transfusion Medicine and Gene Therapy, Medical Center-University of Freiburg, 79106 Freiburg, Germany.
  • Center for Chronic Immunodeficiency (CCI), Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany.
  • San Raffaele Telethon Institute for Gene Therapy, Novel Gene Therapy Strategies Unit, 20132 Milan, Italy.
  • Department of Genomic Medicine, Pfizer-University of Granada-Andalusian Regional Government Centre for Genomics and Oncological Research (GENYO), 18016 Granada, Spain.
  • Center for Non-coding RNA in Technology and Health, Department of Veterinary and Animal Sciences, University of Copenhagen, 1870 Frederiksberg, Denmark.
  • Infection, Immunity, and Inflammation Research and Teaching Department, Great Ormond Street Institute of Child Health, University College London, London WC1N 1EH, UK.
  • Cell and Gene Therapy Safety, Clinical Pharmacology and Safety Sciences R&D, AstraZeneca, Cambridge CB2 0AA, UK.
  • School of Biochemistry and Cell Biology, University College Cork, T12 YN60 Cork, Ireland.

Abstract

Targeted nucleases, primarily CRISPR-Cas-based systems, have revolutionized genome editing by enabling precise modification of target genes or transcripts. Many pre-clinical and clinical studies leverage this technology to develop treatments for human diseases; however, substantial off-target genotoxicity concerns delay its clinical translation. Despite the development of a wide array of tools, assays, and technologies aimed at identifying and quantifying off-target effects, the absence of standardized guidelines leads to inconsistent practices across studies. This review highlights the key challenges and potential solutions in ensuring the safety of gene editing studies for therapeutic applications, focusing on gRNA design, off-target sites prediction, and off-target activity measurement.

MeSH

Genes, Clustered Regularly Interspaced Short Palindromic Repeats, Safety, Genome, Gene Editing, Therapeutics