Reprogramming of cells to a new fate
AssigneeThe Scripps Research Institute
InventorsJem A. Efe, Janghwan Kim, Saiyong Zhu, Simon Hilcove, Sheng Ding
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Abstract
The present invention generally provides methods and compositions for transdifferentiation of an animal cell from a first non-pluripotent cell fate to a second non-pluripotent cell fate. Also provided are methods and compositions for the transdifferentiation of an animal cell from a non-pluripotent mesodermal, endodermal, or ectodermal cell fate to a different non-pluripotent mesodermal, endodermal, or ectodermal cell fate.
Claims29
1-48. (canceled)
A method for screening for small molecules that enhance conversion of an animal cell from a first non-pluripotent cell fate to a second non-pluripotent cell fate, the method comprising:
- (a) introducing a polynucleotide encoding an Oct4 polypeptide, and optionally one or more reprogramming factors comprising polynucleotides encoding a Klf4 polypeptide, a Sox2 polypeptide, or a c-Myc polypeptide, into a first non-pluripotent cell, or
- contacting a first non-pluripotent cell with an Oct4 polypeptide, and optionally one or more reprogramming factors comprising a Klf4 polypeptide, a Sox2 polypeptide, or a c-Myc polypeptide;
- (b) limiting the expression of endogenous Nanog in the cells from step (a) to a level substantially lower than the level of expression of endogenous Nanog in an induced pluripotent cell (iPSC), thereby generating a non-pluripotent intermediate cell;
- (c) contacting the cells from step (b) with a library of different small molecules under conditions to generate a cell having a second non-pluripotent cell fate;
- (d) screening the contacted cells from step (c) for characteristics of a second non-pluripotent cell fate;
- (e) correlating the characteristics of a second non-pluripotent cell fate with a particular small molecule from the library, thereby identifying a small molecule that enhances conversion of an animal cell from a first non-pluripotent cell fate to a second non-pluripotent cell fate.
A composition for efficiently transdifferentiating an animal cell from a first non-pluripotent cell fate to a second non-pluripotent cell fate, the composition comprising:
- (a) a non-pluripotent intermediate cell obtained by a method of
- (i) introducing a polynucleotide encoding an Oct4 polypeptide, and optionally one or more reprogramming factors comprising polynucleotides encoding a Klf polypeptide, a Sox2 polypeptide, or a c-Myc polypeptide;
- or
- contacting cells having a first non-pluripotent cell fate with an Oct4 polypeptide, and optionally one or more reprogramming factors comprising a Klf polypeptide, a Sox2 polypeptide, or a c-Myc polypeptide;
- and
- (ii) limiting the expression of endogenous Nanog in the cells from step (i) to a level substantially lower than the level of expression of endogenous Nanog in an induced pluripotent cell (iPSC), thereby generating a non-pluripotent intermediate cell;
- and
- (b) one or more of a growth factor, a cytokine, and/or a small molecule, wherein the one or more of a growth factor, a cytokine, and/or a small molecular are present in amounts effective to induce differentiation of the non-pluripotent intermediate cell into a cell having a second non-pluripotent cell fate.
Dependencies are inferred from the claim text.