Mutation-independent Allele-Specific Editing by CRISPR-Cas9, a Novel Approach to Treat Autosomal Dominant Disease

Kathleen Christie, Louise Robertson, C Conway, Kevin Blighe, Lawrence Dedionisio, Connie Chao-Shern, Amanda Kowalczyk, John Marshall, Doug Turnbull, M. Andrew Nesbit, Tara C. B. Moore

Research output: Contribution to journalArticlepeer-review

11 Citations (Scopus)
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CRISPR-Cas9 provides a tool to treat autosomal dominant disease by non-homologous end joining (NHEJ) gene disruption of the mutant allele. In order to discriminate between wild-type and mutant alleles, Streptococcus pyogenes Cas9 (SpCas9) must be able to detect a single nucleotide change. Allele-specific editing can be achieved by using either a guide-specific approach, in which the missense mutation is found within the guide sequence, or a protospacer-adjacent motif (PAM)-specific approach, in which the missense mutation generates a novel PAM. While both approaches have been shown to offer allele specificity in certain contexts, in cases where numerous missense mutations are associated with a particular disease, such as TGFBI (transforming growth factor β-induced) corneal dystrophies, it is neither possible nor realistic to target each mutation individually. In this study, we demonstrate allele-specific CRISPR gene editing independent of the disease-causing mutation that is capable of achieving complete allele discrimination, and we propose it as a targeting approach for autosomal dominant disease. Our approach utilizes natural variants in the target region that contain a PAM on one allele that lies in cis with the causative mutation, removing the constraints of a mutation-dependent approach. Our innovative patient-specific guide design approach takes into account the patient’s individual genetic make-up, allowing on- and off-target activity to be assessed in a personalized manner.
Original languageEnglish
Pages (from-to)1846-1857
Number of pages12
JournalMolecular Therapy
Issue number8
Early online date7 May 2020
Publication statusPublished (in print/issue) - 5 Aug 2020


  • CRISPR-Cas9
  • allele specificity
  • autosomal dominant disease
  • gene therapy
  • patient-specific
  • personalised medicine


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