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Gene editing of DNAH11 restores normal cilia motility in primary ciliary dyskinesia
  1. Michele Lai1,
  2. Massimo Pifferi2,
  3. Andrew Bush3,
  4. Martina Piras2,
  5. Angela Michelucci4,
  6. Maria Di Cicco2,
  7. Ambra del Grosso1,
  8. Paola Quaranta1,5,
  9. Chiara Cursi1,
  10. Elena Tantillo1,
  11. Sara Franceschi6,
  12. Maria Chiara Mazzanti6,
  13. Paolo Simi4,
  14. Giuseppe Saggese2,
  15. Attilio Boner7,
  16. Mauro Pistello1
  1. 1Retrovirus Center and Virology Section, Department of Translational Research, University of Pisa, Pisa, Italy
  2. 2Department of Pediatrics, University Hospital of Pisa, Pisa, Italy
  3. 3Imperial College and Royal Brompton Hospital, London, UK
  4. 4Medical Genetics Laboratory, University Hospital of Pisa, Pisa, Italy
  5. 5ARPA Foundation, Pisa, Italy
  6. 6Fondazione Pisana per la Scienza, Pisa, Italy
  7. 7Department of Pediatrics, University of Verona, Verona, Italy
  1. Correspondence to Professor Mauro Pistello, Centro Retrovirus e Sezione Virologia, Dipartimento Ricerca Translazionale e delle Nuove Tecnologie in Medicina e Chirurgia, Università di Pisa, Via San Zeno, 37, Pisa I-56127, Italy; mauro.pistello{at}


Background Primary ciliary dyskinesia (PCD) is a rare autosomal recessive genetic disorder characterised by dysfunction of motile cilia. Ciliary dysmotility causes poor mucociliary clearance and leads to impairment of pulmonary function and severe respiratory infections. PCD has no specific therapy. With the aim to permanently restore gene function and normalise ciliary motility, we used gene editing to replace mutated with wild-type sequence in defective cells.

Methods The target gene was dynein heavy chain 11 (DNAH11), an essential component of ciliary structure. Airway ciliated cells were collected from two patients with PCD with DNAH11 nonsense mutations and altered ciliary beating and pattern. Repair of the genetic defect was performed ex vivo by site-specific recombination using transcription activator-like effector nucleases (TALENs).

Results In an epithelial cell line engineered to contain the DNAH11 target site, TALENs cleaved over 80% of the mutated DNAH11 sequence and replaced the mutated sequence with wild-type sequence in about 50% of cells. In airway ciliated cells of patients with PCD, site-specific recombination and normalisation of ciliary beating and pattern occurred in 33% and 29% of cells, respectively.

Conclusion This study demonstrates that gene editing can rescue ciliary beating ex vivo, opening up new avenues for treating PCD.

  • Primary ciliary diskynesia
  • PCD
  • Gene editing
  • Airway epithelial cells

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