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The transcriptome of the adenovirus infected cell. Zhao, H., Dahlö, M., Isaksson, A., Syvänen, A. Site-specific integration and tailoring of cassette design for sustainable gene transfer. Myelodysplastic syndromes current treatment algorithm 2018. Precise gene editing preserves hematopoietic stem cell function following transient p53-Mediated DNA damage response. High efficiency CRISPR/Cas9-mediated gene editing in primary human T-cells using mutant adenoviral E4orf6/E1b55k ‘helper’ proteins. Increasing the efficiency of homology-directed repair for CRISPR-Cas9-induced precise gene editing in mammalian cells. Advances in genome editing through control of DNA repair pathways. Gene correction for SCID-X1 in long-term hematopoietic stem cells. Site-specific gene editing of human hematopoietic stem cells for X-linked hyper-IgM syndrome. Targeted gene addition in human CD34(+) hematopoietic cells for correction of X-linked chronic granulomatous disease. CRISPR/Cas9 β-globin gene targeting in human haematopoietic stem cells. Homology-driven genome editing in hematopoietic stem and progenitor cells using ZFN mRNA and AAV6 donors. Pyrimidoindole derivatives are agonists of human hematopoietic stem cell self-renewal.
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Aryl hydrocarbon receptor antagonists promote the expansion of human hematopoietic stem cells. Preclinical modeling highlights the therapeutic potential of hematopoietic stem cell gene editing for correction of SCID-X1. Targeted genome editing in human repopulating haematopoietic stem cells. Genome engineering with targetable nucleases. Genetic engineering of hematopoiesis: current stage of clinical translation and future perspectives. This enhanced protocol should broaden applicability of HSC gene editing and pave its way to clinical translation. Combined E4orf6/7 expression and p53 inhibition resulted in HDR editing efficiencies of up to 50% in the long-term human graft, without perturbing repopulation and self-renewal of edited HSCs. We increased HDR efficiency by forcing cell-cycle progression and upregulating components of the HDR machinery through transient expression of the adenovirus 5 E4orf6/7 protein, which recruits the cell-cycle controller E2F on its target genes. Transient p53 inhibition restored polyclonal graft composition. Here, we apply a barcoding strategy to clonal tracking of edited cells (BAR-Seq) and show that editing activates p53, which substantially shrinks the HSC clonal repertoire in hematochimeric mice, although engrafted edited clones preserve multilineage and self-renewing capacity. However, the limited efficiency of homology-directed repair (HDR) in HSCs and the unknown impact of the procedure on clonal composition and dynamics of transplantation have hampered clinical translation. Targeted gene editing in hematopoietic stem cells (HSCs) is a promising treatment for several diseases.