PT  - JOURNAL ARTICLE
AU  - Lai, Michele
AU  - Pifferi, Massimo
AU  - Bush, Andrew
AU  - Piras, Martina
AU  - Michelucci, Angela
AU  - Di Cicco, Maria
AU  - del Grosso, Ambra
AU  - Quaranta, Paola
AU  - Cursi, Chiara
AU  - Tantillo, Elena
AU  - Franceschi, Sara
AU  - Mazzanti, Maria Chiara
AU  - Simi, Paolo
AU  - Saggese, Giuseppe
AU  - Boner, Attilio
AU  - Pistello, Mauro
TI  - Gene editing of <em>DNAH11</em> restores normal cilia motility in primary ciliary dyskinesia
AID  - 10.1136/jmedgenet-2015-103539
DP  - 2016 Apr 01
TA  - Journal of Medical Genetics
PG  - 242--249
VI  - 53
IP  - 4
4099  - http://jmg.bmj.com/content/53/4/242.short
4100  - http://jmg.bmj.com/content/53/4/242.full
SO  - J Med Genet2016 Apr 01; 53
AB  - 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.