Human OKSG-cMyc Simplicon(R) RNA

Various methods utilizing viruses, DNA, RNA, miRNA and protein have been developed to generate integration-free induced pluripotent stem cells (iPSCs). Disadvantages to existing methods include: (1) low reprogramming efficiency (i.e. DNA and protein), (2) a lengthy requirement for negative selection and subcloing steps to remove persistent traces of the virus (i.e. Sendai virus)1 and (3) daily transfections of cells using four synthetic mRNAs over a 14 day period (i.e. mRNA based).EMD Millipore's Simplicon(R) OKSG-cMyc RNA uses a safe and efficient method to generate integration- and virus-free human iPSCs using a single transfection step. The technology utilizes a single self-replicating Venezuelian equine encephalitis (VEE) RNA species that expresses reprogramming factors (RF) ORFs3. The Simplicon RNA replicon is a synthetic polycistronic VEE-RF RNA expressing five reprogramming factors (OKSG-cMyc; Oct4, Klf4, Sox2, Glis1 and cMyc), capable of self-replicating in a limited number of cell divisions. The 5-factor OKSG-cMyc Simplicon is especially useful for iPSCs generation from somatic cells that are more difficult to reprogram (i.e. slower proliferating cells or aged somatic cells).