Method for production of reprogrammed cell using chromosomally unintegrated virus vector
An objective of the present invention is to provide vectors for conveniently and efficiently producing ES-like cells in which foreign genes are not integrated into the chromosome. The present inventors discovered methods for producing ES-like cells from somatic cells using chromosomally non-integrating viral vectors. Since no foreign gene is integrated into the chromosome of the produced ES-like cells, they are advantageous in tests and research, and immunological rejection and ethical problems can be avoided in disease treatments.
1. A Sendai virus vector having a deletion of F gene, and encoding an M protein having mutations of G69E, T116A, and A183S, an HN protein having mutations of A262T, G264R, and K461G, a P protein having mutations of L511F, and a L protein having mutations of N1197S and K1795E, wherein viral protein(s) encoded in said Sendai virus vector further comprises mutations of any one of (i) to (iv) below:
(i) D433A, R434A and K437A in the P protein, and L1361C and L1558I in the L protein
(ii) D433A, R434A and K437A in the P protein;
(iii) Y942H, L1361C and L1558I in the L protein; and
(iv) D433A, R434A and K437A in the P protein, and L1558I in the L protein,
and wherein the Sendai virus vector can be used for expressing reprograming factor genes in a somatic cell to induce an induced pluripotent stem cell.
2. A method for introducing one or more foreign genes into a single cell comprising introducing the Sendai virus vector according to claim 1 into the cell.
3. The method according to claim 2 , further comprising the step of elevating the incubating temperature to remove said vectors.
4. The method according to claim 3 , wherein the incubating temperature is elevated to 37.5° C. to 39° C.
5. A composition for introducing one or more foreign genes into a single cell, comprising a carrier or diluent and the Sendai virus vector according to claim 1 .
6. The composition according to claim 5 , further comprising a Sendai virus vector having a deletion of F gene and encoding an M protein comprising the mutations of G69E, T116A, and A183S, an HN protein comprising the mutations of A262T, G264R, and K461G, a P protein comprising the mutation of L511F, and an L protein comprising the mutations of N1197S and K1795E.