Pavel-Dinu, MaraWiebking, VolkerDejene, Beruh T.Srifa, WarachareeMantri, SruthiNicolas, Carmencita E.Lee, CiaranBao, GangKildebeck, Eric J.Punjya, NirajSindhu, CamilleInlay, Matthew A.Saxena, NiveditaDeRavin, Suk SeeMalech, HarryRoncarolo, Maria GraziaWeinberg, Kenneth I.Porteus, Matthew H.2019-12-122019-12-122019Pavel-Dinu, Mara, Wiebking, Volker, Dejene, Beruh T., et al.. "Gene correction for SCID-X1 in long-term hematopoietic stem cells." <i>Nature Communications,</i> 10, (2019) Springer Nature: https://doi.org/10.1038/s41467-019-09614-y.https://hdl.handle.net/1911/107894Gene correction in human long-term hematopoietic stem cells (LT-HSCs) could be an effective therapy for monogenic diseases of the blood and immune system. Here we describe an approach for X-linked sSevere cCombined iImmunodeficiency (SCID-X1) using targeted integration of a cDNA into the endogenous start codon to functionally correct disease-causing mutations throughout the gene. Using a CRISPR-Cas9/AAV6 based strategy, we achieve up to 20% targeted integration frequencies in LT-HSCs. As measures of the lack of toxicity we observe no evidence of abnormal hematopoiesis following transplantation and no evidence of off-target mutations using a high-fidelity Cas9 as a ribonucleoprotein complex. We achieve high levels of targeting frequencies (median 45%) in CD34+ HSPCs from six SCID-X1 patients and demonstrate rescue of lymphopoietic defect in a patient derived HSPC population in vitro and in vivo. In sum, our study provides specificity, toxicity and efficacy data supportive of clinical development of genome editing to treat SCID-Xl.engThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.Journal articles41467-019-09614-yhttps://doi.org/10.1038/s41467-019-09614-y