US 12,275,950 B2
In vitro method of reprogramming fibroblast cells into induced pluripotent stem cells by transcription factors mRNAs co-transfected with SOCS1 mRNA wherein the mRNAs have a poly-A tail of at least 2000 adenines
Marion Jurk, Pulheim (DE); Stefan Wild, Cologne (DE); and Andreas Bosio, Cologne (DE)
Assigned to MILTENYI BIOTEC B.V. & CO. KG, Bergisch Gladbach (DE)
Appl. No. 16/099,134
Filed by Miltenyi Biotec GmbH, Bergisch Gladbach (DE)
PCT Filed May 2, 2017, PCT No. PCT/EP2017/060345
§ 371(c)(1), (2) Date Nov. 5, 2018,
PCT Pub. No. WO2017/191096, PCT Pub. Date Nov. 9, 2017.
Claims priority of application No. 16168589 (EP), filed on May 6, 2016.
Prior Publication US 2019/0144886 A1, May 16, 2019
Int. Cl. C12N 15/00 (2006.01); C07K 14/47 (2006.01); C12N 5/07 (2010.01); C12N 5/074 (2010.01); C12N 5/0775 (2010.01); C12N 5/0783 (2010.01); C12N 5/0789 (2010.01); C12N 5/10 (2006.01); C12N 15/87 (2006.01)
CPC C12N 15/87 (2013.01) [C07K 14/4703 (2013.01); C12N 5/0636 (2013.01); C12N 5/0662 (2013.01); C12N 5/0696 (2013.01); C12N 5/10 (2013.01)] 5 Claims
 
1. An in vitro method of repeated transfection of a fibroblast cell with at least one mRNA encoding a reprogramming factor, comprising introducing to the fibroblast cell: a) Suppressor Of Cytokine Signaling 1 (SOCS1) mRNA; and simultaneously or subsequently b) the at least one mRNA encoding a reprogramming factor; wherein the step b) is repeated at least once, and wherein the SOCS1 mRNA and the at least one mRNA encoding the reprogramming factor have a poly (A) tail at its 3′ end comprising at least 2000 adenines, respectively, wherein the reprogramming factor induces reprograming of the fibroblast cell into an induced pluripotent stem cell (iPSC), wherein the at least one mRNA encoding the reprogramming factor is an mRNA reprogramming mixture comprising mRNAs encoding for Oct3/4, Sox2, Klf4, Lin28, c-Myc, and Nanog.