NHEJ-Mediated Repair of CRISPR-Cas9-Induced DNA Breaks Efficiently Corrects Mutations in HSPCs from Patients with Fanconi Anemia
| Autor: | Raúl Torres-Ruiz, Marta Corton, Laura Ugalde, Francisco J Roman-Rodriguez, Jordi Surrallés, María José Ramírez, Begoña Díez, Cristina Risueño, Sara Bernal, Massimo Bogliolo, Carmen Ayuso, Juan A. Bueren, Francesca March, Paula Río, Julián Sevilla, Sandra Rodriguez-Perales, Helmut Hanenberg, Lara Álvarez |
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| Rok vydání: | 2019 |
| Předmět: |
DNA End-Joining Repair
Medizin Mice Nude Antigens CD34 Biology Cell Line 03 medical and health sciences Mice 0302 clinical medicine Genome editing INDEL Mutation Fanconi anemia Mice Inbred NOD Genetics Homologous chromosome medicine CRISPR Animals Humans DNA Breaks Double-Stranded Gene Alleles 030304 developmental biology Cell Proliferation Gene Editing 0303 health sciences Fanconi Anemia Complementation Group A Protein Hematopoietic Stem Cell Transplantation High-Throughput Nucleotide Sequencing Cell Biology Genetic Therapy medicine.disease Hematopoietic Stem Cells Cell biology Non-homologous end joining Transplantation Fanconi Anemia Molecular Medicine Stem cell CRISPR-Cas Systems 030217 neurology & neurosurgery |
| Zdroj: | Cell Stem Cell r-IIB SANT PAU. Repositorio Institucional de Producción Científica del Instituto de Investigación Biomédica Sant Pau instname |
| ISSN: | 1934-5909 |
| Popis: | Summary 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. |
| Databáze: | OpenAIRE |
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