Compositions, kits, and methods for making induced pluripotent stem cells using synthetic modified RNAs
AssigneeChildren's Medical Center Corporation
InventorsDerrick Rossi, Luigi Warren
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Abstract
Described herein are synthetic, modified RNAs for changing the phenotype of a cell, such as expressing a polypeptide or altering the developmental potential. Accordingly, provided herein are compositions, methods, and kits comprising synthetic, modified RNAs for changing the phenotype of a cell or cells. These methods, compositions, and kits comprising synthetic, modified RNAs can be used either to express a desired protein in a cell or tissue, or to change the differentiated phenotype of a cell to that of another, desired cell type.
Claims9
A composition for making induced pluripotent stem (iPS) cells comprising synthetic modified ribonucleic acids (RNAs) encoding OCT4, SOX 2, and KLF4, and cell growth medium, wherein each cytosine of each of the synthetic modified RNAs is replaced with 5-methylcytosine and each uracil of each of the synthetic mRNAs is replaced with pseudouracil.
A kit comprising:
- a. a vial comprising a synthetic modified RNA encoding an OCT4 reprogramming factor and a buffer;
- b. a vial comprising a synthetic modified RNA encoding a SOX2 reprogramming factor and a buffer;
- c. a vial comprising a synthetic modified RNA encoding a KLF4 reprogramming factor and a buffer;
- and
- d. packaging and instructions therefor;
- wherein each cytosine of each of the synthetic modified RNAs is replaced with 5-methylcytosine and each uracil of each of the synthetic modified RNAs is replaced with pseudouracil.
A method of reprogramming a somatic fibroblast cell into an induced pluripotent stem (iPS) cell, the method comprising contacting the somatic fibroblast cell with synthetic modified RNAs encoding the reprogramming factors OCT4, SOX 2, and KLF4 at least five consecutive times within 25 days, wherein each cytosine of each of the synthetic modified RNAs is replaced with 5-methylcytosine and each uracil of each of the synthetic modified RNAs is replaced with pseudouracil, thereby producing an induced pluripotent stem (iPS) cell.
Dependencies are inferred from the claim text.