東京大学医科学研究所

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学友会セミナー

学友会セミナー:2017年3月28日

開催日時: 2017年3月28日 17:00 ~ 18:00
開催場所: 1号館2階 2-3会議室
講師: Dong-Sung An
所属: UCLA AIDS Institute/School of Nursing University of California・Professor
演題: Highly Efficient Sendai Virus Mediated CRISPR/Cas9 CCR5 editing in hematopoietic stem cells.
概要:

A HIV cure is possible as demonstrated by the Berlin patient who was cured of
HIV after CCR5-Δ32/Δ32 homozygous deficient bone marrow transplantations. The ease and versatility of the CRISPR/Cas9-mediated gene editing technology has spurred an immense interest in using it to edit the CCR5 gene for anti HIV human hematopoietic stem/progenitor cells (HSPCs) based gene therapy strategies.
To maximize the levels of CCR5 gene modification in HSPCs, we recently developed a thermo sensitive/replication restricted Sendai virus vector for highly efficient CRISPR-Cas9 mediated editing of the CCR5 gene (ts SeV-Cas9-CCR5). Ts SeV-Cas9-CCR5 consistently transduces human fetal liver derived and G-CSF mobilized peripheral blood CD34+HSPCs with high efficiencies (~90%) at a multiplicity of infection of 5, including the CD34+/CD38-/CD90+(Thy1+)/CD49fhigh subpopulation capable of hematopoietic reconstitution by a single cell in a humanized NSG mouse. Remarkably, our SeV-Cas9-CCR5 also edits at CCR5 gene at ~80%. Shifting to 37°C resulted in rapid loss of the SeV vector. Ts SeV-Cas9-CCR5 vector transduced CD34+ HSPCs differentiate into various lineage-specific CFUs by standard in vitro colony forming assays and CD14+/CD33+ macrophages in vitro.
Our current study demonstrated unprecedented efficiency (~80%) of CCR5  gene editing in human HSPCs by a newly developed SeV CRISPR/Cas9 delivery vector. SeV is non-pathogenic in humans, has an established safety record, and has been extensively studied and modified for gene therapy applications. Our SeV-Cas9-CCR5 vector can be developed clinical applications requiring highly efficient gene editing in HSPCs. 

世話人: 〇谷 憲三朗 (ALA先端医療学社会連携研究部門)
 四柳 宏 (感染免疫内科)