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YAC (-globin promoter) forward, 5'-AAACGGTCCCTGGCTAAACT-3', and reverse, 5'- GCTGAAGGGTGCTTCCTTTTT-3'. Bone marrow Lin- cell transplantation. Recipients were 6-8- week-old female C57BL/6 mice (The Jackson laboratory). On the day of transplantation, recipient mice were irradiated (1000 rad). Four hours after irradiation, 1x106 Lin- cells were injected intravenously through the tail vein. This procedure was used for ex vivo HSC transduction studies and for secondary transplantations. The secondary recipients were kept for 16 weeks after transplantation for terminal point analyses. HSC mobilization and in vivo transduction: HSCs were mobilized in mice by subcutaneous (SC) injections of human recombinant G-CSF (250g/kg/mouse/day, 4 days) followed by an SC injection of AMD3100 (5mg/kg) on day 5. In addition, animals received Dexamethasone (10 mg/kg, IP) 16 h and 2 h before virus injection to blunt innate toxicity associated with intravenous HDAd injection. Fourty five minutes after AMD3100, animals were intravenously injected with virus vectors through the retro-orbital plexus (4x1010 viral particles per mouse). In vivo selection: Selection was started at day 2 after transduction. Mice were injected with O6BG (15mg/kg, IP) two times, 30 minutes apart. One hour after the second injection of O6BG, mice were injected (IP) with 5mg/kg BCNU. At days 12 and 26, two more rounds were performed with BCNU doses at 9 and 10mg/kg, respectively. Statistical analyses: Data are presented as means ± standard error of the mean (SEM). For comparisons of multiple groups, one-way and two-way analysis of variance (ANOVA) with Bonferroni post-testing for multiple comparisons was employed. Statistical analysis was performed using GraphPad Prism version 6.01 (GraphPad Software Inc., La Jolla, CA). *p≤0.05, ***p≤0.0001. A P value less than 0.05 was considered significant. Next-generation sequencing data have been deposited to the NCBI Sequence Read Archive (SRA) with accession code PRJNA867400. Animal study approval: All experiments involving animals were conducted in accordance with the institutional guidelines set forth by the University of Washington. The University of Washington is an Association for the Assessment and Accreditation of Laboratory Animal Care International (AALAC)- accredited research institution and all live animal work conducted at this university is in accordance with the Office of Laboratory Animal Welfare (OLAW) Public Health Assurance (PHS) policy, USDA Animal