Selection-free genome editing of the sickle mutation in human adult hematopoietic stem/progenitor cells
| العنوان: | Selection-free genome editing of the sickle mutation in human adult hematopoietic stem/progenitor cells |
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| المؤلفون: | Dana Carroll, Dario Boffelli, Seok Jin Heo, Donald B. Kohn, Tianjiao Wang, Jacob E. Corn, Jennifer R. Berman, Mark A. DeWitt, Mark C. Walters, Therese Mitros, David I. K. Martin, Fabrizia Urbinati, Wendy Magis, Denise P. Muñoz, Nicolas Bray |
| المصدر: | Science Translational Medicine. 8 |
| بيانات النشر: | American Association for the Advancement of Science (AAAS), 2016. |
| سنة النشر: | 2016 |
| مصطلحات موضوعية: | Adult, 0301 basic medicine, Hemoglobin, Sickle, CD34, Anemia, Sickle Cell, Mice, SCID, Biology, Polymorphism, Single Nucleotide, Article, Cell Line, Translational Research, Biomedical, Mice, 03 medical and health sciences, Genome editing, Mice, Inbred NOD, Animals, Humans, Guide RNA, Progenitor cell, Gene, Gene Editing, Mice, Knockout, General Medicine, Hematopoietic Stem Cells, Adult Stem Cells, Haematopoiesis, 030104 developmental biology, Mutation, Immunology, Cancer research, Heterografts, CRISPR-Cas Systems, Stem cell, Ex vivo |
| الوصف: | Genetic diseases of blood cells are prime candidates for treatment through ex vivo gene editing of CD34+ hematopoietic stem/progenitor cells (HSPCs), and a variety of technologies have been proposed to treat these disorders. Sickle Cell Disease (SCD) is a recessive genetic disorder caused by a single nucleotide polymorphism (SNP) in the β-globin gene (HBB). Sickle hemoglobin damages erythrocytes, causing vasoocclusion, severe pain, progressive organ damage, and premature death. Here we optimize design and delivery parameters of a ribonucleoprotein (RNP) complex comprising Cas9 protein and unmodified sgRNA together with a single-stranded DNA oligonucleotide donor (ssODN) to enable efficient replacement of the SCD mutation in human HSPCs. Corrected HSPCs from SCD patients produce less sickle hemoglobin RNA and protein and correspondingly increased wild-type hemoglobin when differentiated into erythroblasts. When engrafted in immunocompromised mice, ex vivo treated human HSPCs maintain SCD gene edits throughout sixteen weeks at a level likely to have clinical benefit. These results demonstrate that an accessible approach combining Cas9 RNP with an ssODN can mediate efficient HSPC genome editing, enables investigator-led exploration of gene editing reagents in primary hematopoietic stem cells, and suggests a path towards the development of new gene editing treatments for SCD and other hematopoietic diseases. |
| تدمد: | 1946-6242 1946-6234 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::0a5540b6e98cf2a7dab8a8557175a050Test https://doi.org/10.1126/scitranslmed.aaf9336Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....0a5540b6e98cf2a7dab8a8557175a050 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 19466242 19466234 |
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