Patent ID: 9249423
Filing Date: 2016-02-02
CPC Classification: A61K,A61P,C12N

Claim Text:
1. A method of re-programming somatic cells comprising transiently transfecting the somatic cells ex vivo with one or more mRNAs in an effective amount to change the somatic cells to a less differentiated state, wherein each of the one or more mRNAs are obtained by in vitro transcription of a linear double-stranded DNA template obtained by Polymerase Chain Reaction (PCR), the DNA template comprising from the 5′ to 3′ direction: an RNA polymerase promoter on the coding strand of the double-stranded DNA, a 5′ untranslated region less than 3,000 nucleotides in length and effective for translation of the mRNAs into detectable polypeptides after transfection into the somatic cells, an open reading frame that encodes the polypeptide, wherein the polypeptide is one that facilitates dedifferentiation of the cell selected from the group consisting of OCT4, SOX2, NANOG, LIN28, KLF4 and MYC, a 3′ untranslated region effective for translation of the mRNAs into detectable polypeptides after transfection into a eukaryotic cell, and a poly(A) stretch of 50-5,000 nucleotides on the coding strand of the double-stranded DNA, wherein the promoter is heterologous to the open reading frame, and wherein the DNA template is not contained within a DNA vector and terminates with the 3′ end of the poly(A) stretch.