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Gene correction for SCID-X1 in long-term hematopoietic stem cells

Pavel-Dinu, Mara, Wiebking, Volker, Dejene, Beruh T., Srifa, Waracharee, Mantri, Sruthi, Nicolas, Carmencita E., Lee, Ciaran, Bao, Gang, Kildebeck, Eric J., Punjya, Niraj, Sindhu, Camille, Inlay, Matthew A., Saxena, Nivedita, DeRavin, Suk See, Malech, Harry, Roncarolo, Maria Grazia, Weinberg, Kenneth I., Porteus, Matthew H.·2019·Nature Communications
DOI 10.1038/s41467-019-09614-yPMC PMC6456568

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Abstract

Gene 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.

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Fig. 1 In vitro, medium scale genome targeting at IL2RG locus. a Diagram of genomic integration and correction outcomes. b Top: schematic of IL2RG corrective donors containing (+tNGFR) or not (–tNGFR) selectable marker. Bottom: IL2RG cDNA targeting frequencies of frozen mobilized peripheral blood CD34+HSPCs (white circles) or freshly purified cord blood male-derived CD34+HSCPs (red circles) derived from medium scale (1.0 × 10 6) genome targeting and measured at day 4. Absolute targeting frequencies measured by ddPCR. Median: 23.2% (+tNGFR, n = 11 biological replicates), median 45% (–tNGFR, n = 13 biological replicates). c Single cell-based methylcellulose assay from mock targeted (nucleofected only) or IL2RG cDNA targeted (–tNGFR donor) CD34+HSPCs. Absolute number of clones are shown (n = 3 biological replicates). d Fraction of the total for each type of colony scored. e Gene correction outcome of SCID-X1 patient 2 derived CD34+HSPCs. Shown is the multi-lineage differentiation using OP9-idll1 in vitro system (n = 23 wells). No growth was derived from uncorrected CD34+cells. LT-HSPCs long-term hematopoietic stem cells, ST-HSC short term hematopoietic stem cells, MPP multi-potent progenitor, CMP common myeloid progenitor, LMPP lymphoid multi-potent progenitor, CLP common lymphoid progenitor, HSPCs hematopoietic stem and progenitor cells, ddPCR droplet digital digital droplet PCR. Mean ± s.e.m.; ns not specific (Welch’s t-test). Source data are available in the Source Data file
Fig. 1 In vitro, medium scale genome targeting at IL2RG locus. a Diagram of genomic integration and correction outcomes. b Top: schematic of IL2RG corrective donors containing (+tNGFR) or not (–tNGFR) selectable marker. Bottom: IL2RG cDNA targeting frequencies of frozen mobilized peripheral blood CD34+HSPCs (white circles) or freshly purified cord blood male-derived CD34+HSCPs (red circles) derived from medium scale (1.0 × 10 6) genome targeting and measured at day 4. Absolute targeting frequencies measured by ddPCR. Median: 23.2% (+tNGFR, n = 11 biological replicates), median 45% (–tNGFR, n = 13 biological replicates). c Single cell-based methylcellulose assay from mock targeted (nucleofected only) or IL2RG cDNA targeted (–tNGFR donor) CD34+HSPCs. Absolute number of clones are shown (n = 3 biological replicates). d Fraction of the total for each type of colony scored. e Gene correction outcome of SCID-X1 patient 2 derived CD34+HSPCs. Shown is the multi-lineage differentiation using OP9-idll1 in vitro system (n = 23 wells). No growth was derived from uncorrected CD34+cells. LT-HSPCs long-term hematopoietic stem cells, ST-HSC short term hematopoietic stem cells, MPP multi-potent progenitor, CMP common myeloid progenitor, LMPP lymphoid multi-potent progenitor, CLP common lymphoid progenitor, HSPCs hematopoietic stem and progenitor cells, ddPCR droplet digital digital droplet PCR. Mean ± s.e.m.; ns not specific (Welch’s t-test). Source data are available in the Source Data fileCC BY · PubMed Central
Fig. 2 Normal hematopoietic reconstitution from IL2RG cDNA targeted CD34+ HSPCs. a Timeline of primary (1°) and secondary (2°) human transplants into sub-lethally irradiated NSG mice. CD34+ HSPCs are derived from umbilical cord blood of healthy male donors. Adult mice transplanted intra-femoral (IF) with either WT CD34+HSPCs (white circles) or mock targeted (yellow circles) or RNP only (black circles) or un-selected IL2RG cDNA targeted (blue-black circles) HSPCs. Three – 4 days old NSG pups transplanted intra-hepatic (IH) with either mock or IL2RG targeted HSPCs. b Combined IF and IH human cells engraftment (hCD45+HLA A-B-C+) 16 weeks after 1° human transplant into indicated organs. c %IL2RG cDNA targeted HSPCs within human graft in indicated organs, quantified by ddPCR. BM (n = 24 mice), SP (n = 24 mice), PB (n = 6 mice) (***p = 0.0008, one-way ANOVA). d Percent human engraftment in indicated organs as in (b) 16 weeks post 2° human CD34+ HSPCs transplant into adult NSG mice. *p-value SP-IH = 0.025, *p-value BM-IF = 0.043 (Welch’s t-test). e %IL2RG targeted HSPCs quantied by ddPCR 32 weeks after engraftment. Median shown. BM bone marrow, SP spleen, PB peripheral blood. Source data are available in the Source Data file
Fig. 2 Normal hematopoietic reconstitution from IL2RG cDNA targeted CD34+ HSPCs. a Timeline of primary (1°) and secondary (2°) human transplants into sub-lethally irradiated NSG mice. CD34+ HSPCs are derived from umbilical cord blood of healthy male donors. Adult mice transplanted intra-femoral (IF) with either WT CD34+HSPCs (white circles) or mock targeted (yellow circles) or RNP only (black circles) or un-selected IL2RG cDNA targeted (blue-black circles) HSPCs. Three – 4 days old NSG pups transplanted intra-hepatic (IH) with either mock or IL2RG targeted HSPCs. b Combined IF and IH human cells engraftment (hCD45+HLA A-B-C+) 16 weeks after 1° human transplant into indicated organs. c %IL2RG cDNA targeted HSPCs within human graft in indicated organs, quantified by ddPCR. BM (n = 24 mice), SP (n = 24 mice), PB (n = 6 mice) (***p = 0.0008, one-way ANOVA). d Percent human engraftment in indicated organs as in (b) 16 weeks post 2° human CD34+ HSPCs transplant into adult NSG mice. *p-value SP-IH = 0.025, *p-value BM-IF = 0.043 (Welch’s t-test). e %IL2RG targeted HSPCs quantied by ddPCR 32 weeks after engraftment. Median shown. BM bone marrow, SP spleen, PB peripheral blood. Source data are available in the Source Data fileCC BY · PubMed Central
Fig. 3 Normal multi-lineage development from IL2RG cDNA targeted in the LT-HSC population. a Percent cellular composition of the lymphoid, myeloid and erythroid lineage derived from IH 1° human engraftment, shown in indicated organs and targeting conditions. CD3+ BM: **p = 0.0017, CD3+SP: **p = 0.007 (Welch’s t-test). b Same as (a) but IF transplant analysis. CD3+ SP: *p = 0.023, CD56+ BM: *p = 0.015 (Kruskal–Wallis test). c Percent cellular composition of the lymphoid, myeloid and erythroid lineage derived from secondary transplants. Data shown are combined IH and IF primary transplants. CD3+ BM: *p = 0.015, CD56+: *p = 0.025, CD19+ SP: ***p = 0.0002, CD14+ SP: *p = 0.0112, CD11c+ SP: ***p = 0.0004. LT long term. Error bars: mean ± s.e.m. Source data are available in the Source Data file
Fig. 3 Normal multi-lineage development from IL2RG cDNA targeted in the LT-HSC population. a Percent cellular composition of the lymphoid, myeloid and erythroid lineage derived from IH 1° human engraftment, shown in indicated organs and targeting conditions. CD3+ BM: **p = 0.0017, CD3+SP: **p = 0.007 (Welch’s t-test). b Same as (a) but IF transplant analysis. CD3+ SP: *p = 0.023, CD56+ BM: *p = 0.015 (Kruskal–Wallis test). c Percent cellular composition of the lymphoid, myeloid and erythroid lineage derived from secondary transplants. Data shown are combined IH and IF primary transplants. CD3+ BM: *p = 0.015, CD56+: *p = 0.025, CD19+ SP: ***p = 0.0002, CD14+ SP: *p = 0.0112, CD11c+ SP: ***p = 0.0004. LT long term. Error bars: mean ± s.e.m. Source data are available in the Source Data fileCC BY · PubMed Central
Fig. 4 In vivo rescue of SCID-X1 mutation. a Genomic mapping and description of SCID-X1 mutations. b Percent viability determined at indicated days pre- and post- targeting. Mock (nucleofected only), RNP (nucleofected with RNP only), RNP+AAV6 (nucleofected with RNP and transduced with AAV6-based IL2RG corrective donor). Shown is data for mobilized peripheral blood CD34+ HSPCs (n = 5). c Medium scale (1.0 × 106 cells) ex vivo genome targeting frequencies of frozen mobilized peripheral blood SCID-X1, at day 2 (blue-black circles, n = 6). Arrow shows 45% genome targeting of SCID-X1 patient 2 derived CD34+ HSPCs. d Human cells engraftment analysis at week 17 after intra-hepatic (IH) delivery of IL2RG cDNA targeted (blue-black circles, n = 15) or mutant CD34+ HSPCs (gray circles, n = 4). e Percent cellular composition of the lymphoid, myeloid, and erythroid lineage derived from IL2RG corrected or mutant CD34+ HSPCs. CD3+: ****p < 0.0001, CD56+: *p = 0.0146, CD16+: **p = 0.0013, CD19+: **p = 0.0015, CD235a+: **p = 0.0022 (Welch’s t-test). RNP ribonuclearprotein. f Absolute numbers derived from (e). Source data are available in the Source Data file
Fig. 4 In vivo rescue of SCID-X1 mutation. a Genomic mapping and description of SCID-X1 mutations. b Percent viability determined at indicated days pre- and post- targeting. Mock (nucleofected only), RNP (nucleofected with RNP only), RNP+AAV6 (nucleofected with RNP and transduced with AAV6-based IL2RG corrective donor). Shown is data for mobilized peripheral blood CD34+ HSPCs (n = 5). c Medium scale (1.0 × 106 cells) ex vivo genome targeting frequencies of frozen mobilized peripheral blood SCID-X1, at day 2 (blue-black circles, n = 6). Arrow shows 45% genome targeting of SCID-X1 patient 2 derived CD34+ HSPCs. d Human cells engraftment analysis at week 17 after intra-hepatic (IH) delivery of IL2RG cDNA targeted (blue-black circles, n = 15) or mutant CD34+ HSPCs (gray circles, n = 4). e Percent cellular composition of the lymphoid, myeloid, and erythroid lineage derived from IL2RG corrected or mutant CD34+ HSPCs. CD3+: ****p < 0.0001, CD56+: *p = 0.0146, CD16+: **p = 0.0013, CD19+: **p = 0.0015, CD235a+: **p = 0.0022 (Welch’s t-test). RNP ribonuclearprotein. f Absolute numbers derived from (e). Source data are available in the Source Data fileCC BY · PubMed Central
Fig. 5 Evaluation of IL-2 receptor function in IL2RG cDNA targeted T cells. a Schematic of signaling (pSTAT5—bottom) and proliferation (CFSE—top) in vitro assays. b pSTAT5 assay derived FACS plots. Top: healthy male-derived T cells genome targeted with IL2RG cDNA tNGFR (KI) or with tNGFR+ only cassette integrated at the IL2RG endogenous locus (KO). In red are the percent of double positive IL2RG-tNGFR+pSTAT5+[4.42%/(4/42% + 3.18%)]×100. We compare 58.2% cells (IL2RG targeted T cells) with 58.7% (IL2RG from WT T cells), (n = 3 biological replicates). c Quantification of IL-2R signaling through phosphoSTAT5 pathway. d pSTAT5 MFI for WT, KI, and KO experiments from (b) p = 0.02, Welch’s t-test. WT T cells (gray circles, n = 6), IL2RG KI (blue circles, n = 3) and IL2RG KO (orange circles, n = 3). e Proliferation profile of CFSE labeled, TCR stimulated IL2RG cDNA tNGFR+ sorted or mock-targeted T cells. Mock-targeted T cells are WT T cells cultured for the same amount of time as the tNGFR+ targeted cells and have been nucleofected in the absence of RNP or absence of transduction with AAV6. Shown FACS analysis at days 2, 4, 6, and 8. pSTAT5 phosphorylated STAT5, CFSE carboxyfluorescein succinimidyl ester, KI knocked in, KO knocked out, tNGFR truncated nerve growth factor receptor, IL-2 interleukin 2. Source data are available in the Source Data file
Fig. 5 Evaluation of IL-2 receptor function in IL2RG cDNA targeted T cells. a Schematic of signaling (pSTAT5—bottom) and proliferation (CFSE—top) in vitro assays. b pSTAT5 assay derived FACS plots. Top: healthy male-derived T cells genome targeted with IL2RG cDNA tNGFR (KI) or with tNGFR+ only cassette integrated at the IL2RG endogenous locus (KO). In red are the percent of double positive IL2RG-tNGFR+pSTAT5+[4.42%/(4/42% + 3.18%)]×100. We compare 58.2% cells (IL2RG targeted T cells) with 58.7% (IL2RG from WT T cells), (n = 3 biological replicates). c Quantification of IL-2R signaling through phosphoSTAT5 pathway. d pSTAT5 MFI for WT, KI, and KO experiments from (b) p = 0.02, Welch’s t-test. WT T cells (gray circles, n = 6), IL2RG KI (blue circles, n = 3) and IL2RG KO (orange circles, n = 3). e Proliferation profile of CFSE labeled, TCR stimulated IL2RG cDNA tNGFR+ sorted or mock-targeted T cells. Mock-targeted T cells are WT T cells cultured for the same amount of time as the tNGFR+ targeted cells and have been nucleofected in the absence of RNP or absence of transduction with AAV6. Shown FACS analysis at days 2, 4, 6, and 8. pSTAT5 phosphorylated STAT5, CFSE carboxyfluorescein succinimidyl ester, KI knocked in, KO knocked out, tNGFR truncated nerve growth factor receptor, IL-2 interleukin 2. Source data are available in the Source Data fileCC BY · PubMed Central
Fig. 6 Genome specificity of IL2RG sgRNA guide. a Heat map of on-target INDEL frequencies quantied by NexGen-Seq at COSMID identified putative on-target locations from healthy CD34+ HSPCs. Levels of NHEJ induced by 20 nt IL2RG sgRNA and truncated 19 nt, 18 nt and 17 nt pre-complexed with WT Cas9 protein at 5:1 molar ratio. b Heat map as in (a) of on-target INDEL frequencies derived from 19 nt IL2RG sg-1 in the genome of CD34+ HSPCs SCID-X1 patient 1 derived cells. c Percent viability at day 4 of SCID-X1 patient-derived CD34+ HSPCs nucleofected with either wild-type (WT) or high-fidelity (HiFi) SpCas9 protein pre-complexed with either the 20 nt or the 19 nt IL2RG sg-1 (n = 1). d Percent INDELs measured by TIDE at day 4 in cells as in (c) using WT or HiFi Cas9 protein pre-complexed with the 20 nt IL2RG sg-1 (green bars) or 19 nt IL2RG sg-1 (blue bars). e Percent IL2RG cDNA targeting (% HR) as measured by ddPCR at day 4 in cells as in (c) generated by either WT or HiFi Cas9 protein pre-complexed with the 20 nt IL2RG sg-1 or (f) 19 nt IL2RG sg-1. Source data are available in the Source Data file
Fig. 6 Genome specificity of IL2RG sgRNA guide. a Heat map of on-target INDEL frequencies quantied by NexGen-Seq at COSMID identified putative on-target locations from healthy CD34+ HSPCs. Levels of NHEJ induced by 20 nt IL2RG sgRNA and truncated 19 nt, 18 nt and 17 nt pre-complexed with WT Cas9 protein at 5:1 molar ratio. b Heat map as in (a) of on-target INDEL frequencies derived from 19 nt IL2RG sg-1 in the genome of CD34+ HSPCs SCID-X1 patient 1 derived cells. c Percent viability at day 4 of SCID-X1 patient-derived CD34+ HSPCs nucleofected with either wild-type (WT) or high-fidelity (HiFi) SpCas9 protein pre-complexed with either the 20 nt or the 19 nt IL2RG sg-1 (n = 1). d Percent INDELs measured by TIDE at day 4 in cells as in (c) using WT or HiFi Cas9 protein pre-complexed with the 20 nt IL2RG sg-1 (green bars) or 19 nt IL2RG sg-1 (blue bars). e Percent IL2RG cDNA targeting (% HR) as measured by ddPCR at day 4 in cells as in (c) generated by either WT or HiFi Cas9 protein pre-complexed with the 20 nt IL2RG sg-1 or (f) 19 nt IL2RG sg-1. Source data are available in the Source Data fileCC BY · PubMed Central

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Research works printed but not linked: 41

  1. Pai, S. Y. et al. Transplantation outcomes for severe combined immunodeficiency, 2000-2009. N. Engl. J. Med. 371, 434–446 (2014). · DOI
  2. De Ravin, S. S. et al. Lentiviral hematopoietic stem cell gene therapy for X-linked severe combined immunodeficiency. Sci. Transl. Med. 8, 335ra357 (2016). · DOI · Paper page
  3. Stephan, V. et al. Atypical X-linked severe combined immunodeficiency due to possible spontaneous reversion of the genetic defect in T cells. N. Engl. J. Med. 335, 1563–1567 (1996). · DOI
  4. Hacein-Bey-Abina, S. et al. LMO2-associated clonal T cell proliferation in two patients after gene therapy for SCID-X1. Science 302, 415–419 (2003). · DOI · Paper page
  5. Woods, N. B., Bottero, V., Schmidt, M., von Kalle, C. & Verma, I. M. Gene therapy: therapeutic gene causing lymphoma. Nature 440, 1123 (2006). · DOI
  6. Aiuti, A. & Roncarolo, M. G. Ten years of gene therapy for primary immune deficiencies. Hematol. Am. Soc. Hematol. Educ. Prog. https://doi.org/10.1182/asheducation-2009.1.682 (2009). · DOI
  7. Fischer, A., Hacein-Bey-Abina, S. & Cavazzana-Calvo, M. 20 years of gene therapy for SCID. Nat. Immunol. 11, 457–460 (2010). · DOI
  8. Smogorzewska, E. M., Weinberg, K. I. & Kohn, D. B. [Transplantation of genetically modified cells in the treatment of children with SCID: great hopes and recent disappointments]. Med. Wieku. Rozwoj. 7, 27–34 (2003).
  9. Ott, M. G. et al. Correction of X-linked chronic granulomatous disease by gene therapy, augmented by insertional activation of MDS1-EVI1, PRDM16 or SETBP1. Nat. Med. 12, 401–409 (2006). · DOI · Paper page
  10. Kang, E. M. et al. Retrovirus gene therapy for X-linked chronic granulomatous disease can achieve stable long-term correction of oxidase activity in peripheral blood neutrophils. Blood 115, 783–791 (2010). · DOI
  11. Boztug, K. et al. Stem-cell gene therapy for the Wiskott-Aldrich syndrome. N. Engl. J. Med. 363, 1918–1927 (2010). · DOI · Paper page
  12. Cavazzana, M., Six, E., Lagresle-Peyrou, C., Andre-Schmutz, I. & Hacein-Bey-Abina, S. Gene therapy for X-linked severe combined immunodeficiency: where do we stand? Hum. Gene Ther. 27, 108–116 (2016). · DOI · Paper page
  13. Thornhill, S. I. et al. Self-inactivating gammaretroviral vectors for gene therapy of X-linked severe combined immunodeficiency. Mol. Ther. 16, 590–598 (2008). · DOI
  14. Voit, R. A., Hendel, A., Pruett-Miller, S. M. & Porteus, M. H. Nuclease-mediated gene editing by homologous recombination of the human globin locus. Nucleic Acids Res. 42, 1365–1378 (2014). · DOI
  15. Schiroli, G. et al. Preclinical modeling highlights the therapeutic potential of hematopoietic stem cell gene editing for correction of SCID-X1. Sci. Transl. Med. https://doi.org/10.1126/scitranslmed.aan0820 (2017). · DOI · Paper page
  16. Porteus, M. H., Cathomen, T., Weitzman, M. D. & Baltimore, D. Efficient gene targeting mediated by adeno-associated virus and DNA double-strand breaks. Mol. Cell. Biol. 23, 3558–3565 (2003). · DOI
  17. DiGiusto, D. L. et al. Preclinical development and qualification of ZFN-mediated CCR5 disruption in human hematopoietic stem/progenitor cells. Mol. Ther. Methods Clin. Dev. 3, 16067 (2016). · DOI
  18. Urnov, F. D. et al. Highly efficient endogenous human gene correction using designed zinc-finger nucleases. Nature 435, 646–651 (2005). · DOI
  19. Connelly, J. P., Barker, J. C., Pruett-Miller, S. & Porteus, M. H. Gene correction by homologous recombination with zinc finger nucleases in primary cells from a mouse model of a generic recessive genetic disease. Mol. Ther. 18, 1103–1110 (2010). · DOI
  20. Li, T. et al. TAL nucleases (TALNs): hybrid proteins composed of TAL effectors and FokI DNA-cleavage domain. Nucleic Acids Res. 39, 359–372 (2011). · DOI
  21. Jinek, M. et al. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science 337, 816–821 (2012). · DOI · Paper page
  22. Hendel, A. et al. Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells. Nat. Biotechnol. 33, 985–989 (2015). · DOI · Paper page
  23. Sentmanat, M. F., Peters, S. T., Florian, C. P., Connelly, J. P. & Pruett-Miller, S. M. A survey of validation strategies for CRISPR-Cas9 editing. Sci. Rep. 8, 888 (2018). · DOI
  24. Inlay, M. A. et al. Identification of multipotent progenitors that emerge prior to hematopoietic stem cells in embryonic development. Stem Cell. Rep. 2, 457–472 (2014). · DOI
  25. Hong, C., Luckey, M. A. & Park, J. H. Intrathymic IL-7: the where, when, and why of IL-7 signaling during T cell development. Semin. Immunol. 24, 151–158 (2012). · DOI
  26. Seet, C. S. et al. Generation of mature T cells from human hematopoietic stem and progenitor cells in artificial thymic organoids. Nat. Methods 14, 521–530 (2017). · DOI
  27. Lu, P. H. & Negrin, R. S. A novel population of expanded human CD3+CD56+cells derived from T cells with potent in vivo antitumor activity in mice with severe combined immunodeficiency. J. Immunol. 153, 1687–1696 (1994). · DOI
  28. Traggiai, E. et al. Development of a human adaptive immune system in cord blood cell-transplanted mice. Science 304, 104–107 (2004). · DOI
  29. Orr, S. J. et al. Implications for gene therapy-limiting expression of IL-2R gamma c delineate differences in signaling thresholds required for lymphocyte development and maintenance. J. Immunol. 185, 1393–1403 (2010). · DOI
  30. Cradick, T. J., Qiu, P., Lee, C. M., Fine, E. J. & Bao, G. COSMID: a web-based tool for identifying and validating CRISPR/Cas off-target sites. Mol. Ther. Nucleic Acids 3, e214 (2014). · DOI · Paper page
  31. Holt, N. et al. Human hematopoietic stem/progenitor cells modified by zinc-finger nucleases targeted to CCR5 control HIV-1 in vivo. Nat. Biotechnol. 28, 839–847 (2010). · DOI · Paper page
  32. Hendel, A. et al. Quantifying genome-editing outcomes at endogenous loci with SMRT sequencing. Cell Rep. 7, 293–305 (2014). · DOI
  33. Pruett-Miller, S. M., Reading, D. W., Porter, S. N. & Porteus, M. H. Attenuation of zinc finger nuclease toxicity by small-molecule regulation of protein levels. PLoS. Genet. 5, e1000376 (2009). · DOI
  34. Menon, T. et al. Lymphoid regeneration from gene-corrected SCID-X1 subject-derived iPSCs. Cell. Stem. Cell. 16, 367–372 (2015). · DOI
  35. Matsubara, Y. et al. Transcription activator-like effector nuclease-mediated transduction of exogenous gene into IL2RG locus. Sci. Rep. 4, 5043 (2014). · DOI
  36. Hoban, M. D. et al. Correction of the sickle cell disease mutation in human hematopoietic stem/progenitor cells. Blood 125, 2597–2604 (2015). · DOI · Paper page
  37. Ishikawa, F. et al. Development of functional human blood and immune systems in NOD/SCID/IL2 receptor {gamma} chain(null) mice. Blood 106, 1565–1573 (2005). · DOI
  38. McDermott, S. P., Eppert, K., Lechman, E. R., Doedens, M. & Dick, J. E. Comparison of human cord blood engraftment between immunocompromised mouse strains. Blood 116, 193–200 (2010). · DOI
  39. Denning, S. M., Tuck, D. T., Singer, K. H. & Haynes, B. F. Human thymic epithelial cells function as accessory cells for autologous mature thymocyte activation. J. Immunol. 138, 680–686 (1987). · DOI
  40. Cong, L. et al. Multiplex genome engineering using CRISPR/Cas systems. Science 339, 819–823 (2013). · DOI · Paper page
  41. Lee, C. M., Cradick, T. J. & Bao, G. The Neisseria meningitidis CRISPR-Cas9 system enables specific genome editing in mammalian cells. Mol. Ther. 24, 645–654 (2016). · DOI

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Authors
Pavel-Dinu, M., Wiebking, V., Dejene, B. T., Srifa, W., Mantri, S., Nicolas, C. E., Lee, C., Bao, G., Kildebeck, E. J., Punjya, N., Sindhu, C., Inlay, M. A., Saxena, N., DeRavin, S. S., Malech, H., Roncarolo, M. G., Weinberg, K. I., & Porteus, M. H.
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Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2019-04-09
Volume
10
Issue
1
ISSN
2041-1723
DOI
10.1038/s41467-019-09614-y
PMC
PMC6456568
Cited by
167
References
16
Language
en
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Pavel-Dinu, M., Wiebking, V., Dejene, B. T., Srifa, W., Mantri, S., Nicolas, C. E., Lee, C., Bao, G., Kildebeck, E. J., Punjya, N., Sindhu, C., Inlay, M. A., Saxena, N., DeRavin, S. S., Malech, H., Roncarolo, M. G., Weinberg, K. I., & Porteus, M. H. (2019). Gene correction for SCID-X1 in long-term hematopoietic stem cells. Nature Communications. https://doi.org/10.1038/s41467-019-09614-y