Constructs for making induced pluripotent stem cells
Systems, constructs, and methods for reprogramming cells are provided. In one aspect, for example, a transformation construct for generating iPS cells can include an expression vector having a plurality of reprogramming factors, each reprogramming factor being under control of a separate promoter.
1. A transformation construct for generating induced pluripotent stem (iPS) cells, comprising:
an expression vector including a plurality of reprogramming factors, each reprogramming factor being under control of a separate promoter, wherein the plurality of reprogramming factors includes OCT3/4, SOX2, and at least one member selected from the group consisting of KLF4, c-Myc, NANOG, and LIN28.
2. The construct of claim 1 , wherein the expression vector is selected from the group consisting of plasmids, viruses, and combinations thereof.
3. The construct of claim 1 , wherein the expression vector is selected from the group consisting of adenoviral vectors, episomal vectors, retroviral vectors, and lentiviral vectors.
4. The construct of claim 1 , wherein the expression vector is an episomal vector.
5. The construct of claim 1 , wherein the plurality of reprogramming factors includes OCT3/4, SOX2, KLF4, and c-Myc.
6. The construct of claim 1 , wherein the plurality of reprogramming factors consists of OCT3/4, SOX2, and KLF4.
7. The construct of claim 1 , wherein the plurality of reprogramming factors includes OCT3/4, SOX2, NANOG, and LIN28.
8. The construct of claim 1 , wherein the expression vector has a sequence that is at least 80% homologous to SEQ ID 72.
9. The construct of claim 1 , wherein the expression vector has a sequence that is at least 95% homologous to SEQ ID 72.
10. The construct of claim 1 , wherein at least one of reprogramming factor is under the control of a CMV promoter.
11. The construct of claim 10 , wherein the CMV promoter is a weak CMV promoter.
12. The construct of claim 1 , wherein the expression vector further includes a reporter sequence under control of a separate promoter.