Efficient method for nuclear reprogramming
First Claim
1. A method of preparing an induced pluripotent stem cell from a mammalian somatic cell, comprising nuclear reprogramming the mammalian somatic cell under culture conditions which allow for mammalian somatic cell reprogramming to pluripotency with at least one miRNA and an expression vector encoding at least one nuclear reprogramming factor,wherein the at least one miRNA increases efficiency of the nuclear reprogramming of the mammalian somatic cell compared to nuclear reprogramming of the mammalian somatic cell with the expression vector in the absence of the at least one miRNA,wherein the at least one miRNA is one or two or more miRNAs contained in one or two or more RNAs selected from the group consisting of:
- hsa-miR-372;
hsa-miR-373 or hsa-miR-373/373*;
hsa-miR-371-373 cluster;
hsa-miR-302b or hsa-miR-302b/302b*;
hsa-miR-302-367 cluster;
hsa-miR-520c or hsa-miR-520c-5p/520c-3p;
mmu-miR-291a or mmu-miR-291a-5p/291a-3p;
mmu-miR-294 or mmu-miR-294/294*; and
mmu-miR-295 or mmu-miR-295/295*, andwherein the at least one nuclear reprogramming factor comprises at least (i) an Oct3/4 gene, (ii) an Oct3/4 gene and a Klf4 gene, (iii) an Oct3/4 gene and a Nanog gene, or (iv) an Oct3/4 gene, Klf4 gene, and a Myc family gene, but it does not comprise a Sox family gene.
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Abstract
This relates to a method of preparing induced pluripotent stem cells, comprising a nuclear reprogramming step with a nuclear reprogramming factor in the presence of miRNA, wherein said miRNA has a property of providing a higher nuclear reprogramming efficiency in the presence of said miRNA than in the absence thereof.
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Citations
9 Claims
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1. A method of preparing an induced pluripotent stem cell from a mammalian somatic cell, comprising nuclear reprogramming the mammalian somatic cell under culture conditions which allow for mammalian somatic cell reprogramming to pluripotency with at least one miRNA and an expression vector encoding at least one nuclear reprogramming factor,
wherein the at least one miRNA increases efficiency of the nuclear reprogramming of the mammalian somatic cell compared to nuclear reprogramming of the mammalian somatic cell with the expression vector in the absence of the at least one miRNA, wherein the at least one miRNA is one or two or more miRNAs contained in one or two or more RNAs selected from the group consisting of: - hsa-miR-372;
hsa-miR-373 or hsa-miR-373/373*;
hsa-miR-371-373 cluster;
hsa-miR-302b or hsa-miR-302b/302b*;
hsa-miR-302-367 cluster;
hsa-miR-520c or hsa-miR-520c-5p/520c-3p;
mmu-miR-291a or mmu-miR-291a-5p/291a-3p;
mmu-miR-294 or mmu-miR-294/294*; and
mmu-miR-295 or mmu-miR-295/295*, andwherein the at least one nuclear reprogramming factor comprises at least (i) an Oct3/4 gene, (ii) an Oct3/4 gene and a Klf4 gene, (iii) an Oct3/4 gene and a Nanog gene, or (iv) an Oct3/4 gene, Klf4 gene, and a Myc family gene, but it does not comprise a Sox family gene. - View Dependent Claims (3, 4, 5, 6, 7, 8, 9)
- hsa-miR-372;
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2. A method of increasing the efficiency of nuclear reprogramming for inducing an induced pluripotent stem cell from a mammalian somatic cell, comprising
subjecting the mammalian somatic cell to nuclear reprogramming with an expression vector encoding at least one nuclear reprogramming factor in the presence of at least one miRNA under culture conditions which allow for mammalian somatic cell reprogramming to pluripotency, wherein the at least one miRNA increases efficiency of the nuclear reprogramming of the mammalian somatic cell compared to nuclear reprogramming of the somatic cell with the expression vector in the absence of the at least one miRNA, wherein the at least one miRNA is one or two or more miRNAs contained in one or two or more RNAs selected from the group consisting of: - hsa-miR-372;
hsa-miR-373 or hsa-miR-373/373*;
hsa-miR-371-373 cluster;
hsa-miR-302b or hsa-miR-302b/302b*;
hsa-miR-302-367 cluster;
hsa-miR-520c or hsa-miR-520c-5p/520c-3p;
mmu-miR-291a or mmu-miR-291a-5p/291a-3p;
mmu-miR-294 or mmu-miR-294/294*; and
mmu-miR-295 or mmu-miR-295/295*, andwherein the at least one nuclear reprogramming factor comprises at least (i) an Oct3/4 gene, (ii) an Oct3/4 gene and a Klf4 gene, (iii) an Oct3/4 gene and a Nanog gene, or (iv) an Oct3/4 gene, a Klf4 gene, and a Myc family gene, but it does not comprise a Sox family gene.
- hsa-miR-372;
Specification