Selection-free genome editing of the sickle mutation in human adult hematopoietic stem/progenitor cells.

ABSTRACT: Genetic diseases of blood cells are prime candidates for treatment through ex vivo gene editing of CD34⁺ hematopoietic stem/progenitor cells (HSPCs), and a variety of technologies have been proposed to treat these disorders. Sickle cell disease (SCD) is a recessive genetic disorder caused by a single-nucleotide polymorphism in the β-globin gene (HBB). Sickle hemoglobin damages erythrocytes, causing vasoocclusion, severe pain, progressive organ damage, and premature death. We optimize design and delivery parameters of a ribonucleoprotein (RNP) complex comprising Cas9 protein and unmodified single guide RNA, together with a single-stranded DNA oligonucleotide donor (ssODN), to enable efficient replacement of the SCD mutation in human HSPCs. Corrected HSPCs from SCD patients produc

SUBMITTER: DeWitt MA 

PROVIDER: S-EPMC5500303 | biostudies-literature | 2016 Oct

REPOSITORIES: biostudies-literature