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

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

SUBMITTER: Pavel-Dinu M 

PROVIDER: S-EPMC6456568 | biostudies-literature | 2019 Apr

REPOSITORIES: biostudies-literature