NHEJ-Mediated Repair of CRISPR-Cas9-Induced DNA Breaks Efficiently Corrects Mutations in HSPCs from Patients with Fanconi Anemia

Francisco José Román-Rodríguez, Laura Ugalde, Lara Álvarez, Begoña Díez, María José Ramírez, Cristina Risueño, Marta Cortón, Massimo Bogliolo, Sara Bernal, Francesca March, Carmen Ayuso, Helmut Hanenberg, Julián Sevilla, Sandra Rodríguez-Perales, Raúl Torres-Ruiz, Jordi Surrallés, Juan Antonio Bueren, Paula Río*

*Autor corresponent d’aquest treball

Producció científica: Contribució a una revistaArticleRecercaAvaluat per experts

57 Cites (Scopus)

Resum

Non-homologous end-joining (NHEJ) is the preferred mechanism used by hematopoietic stem cells (HSCs) to repair double-stranded DNA breaks and is particularly increased in cells deficient in the Fanconi anemia (FA) pathway. Here, we show feasible correction of compromised functional phenotypes in hematopoietic cells from multiple FA complementation groups, including FA-A, FA-C, FA-D1, and FA-D2. NHEJ-mediated repair of targeted CRISPR-Cas9-induced DNA breaks generated compensatory insertions and deletions that restore the coding frame of the mutated gene. NHEJ-mediated editing efficacy was initially verified in FA lymphoblastic cell lines and then in primary FA patient-derived CD34+ cells, which showed marked proliferative advantage and phenotypic correction both in vitro and after transplantation. Importantly, and in contrast to homologous directed repair, NHEJ efficiently targeted primitive human HSCs, indicating that NHEJ editing approaches may constitute a sound alternative for editing self-renewing human HSCs and consequently for treatment of FA and other monogenic diseases affecting the hematopoietic system.

Idioma originalEnglish
Pàgines (de-a)607-621.e7
Nombre de pàgines22
RevistaCell Stem Cell
Volum25
Número5
DOIs
Estat de la publicacióPublicada - 7 de nov. 2019

Fingerprint

Navegar pels temes de recerca de 'NHEJ-Mediated Repair of CRISPR-Cas9-Induced DNA Breaks Efficiently Corrects Mutations in HSPCs from Patients with Fanconi Anemia'. Junts formen un fingerprint únic.

Com citar-ho