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AssigneeNational Institute of Advanced Industrial Science and Technology
InventorsMahito Nakanishi, Ken Nishimura, Manami Ohtaka, Masayuki Sano
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Stem cell reprogramming genes cloned into a single sustained expression-type Sendai viral vector are shown to reprogram differentiated somatic cells into induced pluripotent stem (iPS) cells without integration of vector sequences into the host cell's genome. The genes are transduced into normal differentiated somatic cells via infection with recombinant Sendai virus. After expression of the reprogramming genes and subsequent induction of pluripotency, the vector genome RNA including the reprogramming genes is removed from the cell to establish an iPS cell that is genetically identical to the parent somatic differentiated cell thus reducing the risk of tumorigenic transformation caused by random integration of vector sequences into the host genome. The method promises to provide safe, autologous iPS cells that can be used for human cell replacement and regeneration therapeutic applications.
A reprogramming gene-loaded Sendai viral vector comprising Sendai virus genes and reprogramming genes comprising a combination of Oct3/4, Sox2 and Klf4, or a combination of Oct3/4, Sox2, Klf4 and c-Myc, wherein the Sendai virus genes include an NP gene, P/C gene, M gene, F gene, HN gene and L gene, wherein each of the M gene, the F gene and the HN gene is from a Sendai virus strain C1.151-derived gene and wherein at least one of the M gene, the F gene and the HN gene is functionally deleted and the L gene encodes the amino-acid sequence of the L protein in which the amino-acid residue at position 1618 is valine.
A template vector for preparing a reprogramming gene-loaded Sendai virus, the template vector comprising a cloning vector with Sendai virus genes and reprogramming genes comprising a combination of Oct3/4, Sox2 and Klf4, or a combination of Oct3/4, Sox2, Klf4 and c-Myc, wherein the Sendai virus genes include an NP gene, P/C gene, M gene, F gene, HN gene and L gene, and wherein each of the M gene, the F gene and the HN gene is from the Sendai virus strain C1.151-derived gene and at least one of respective functions of the M gene, the F gene and the HN gene is functionally deleted;
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