Episome Reprogramming

We offer two options for integration free episome reprogramming:

The Epi5 method is intended for peripheral blood mononuclear cell (PBMC) reprogramming with very low probability for plasmid integration. The protocol includes an option for enriching the CD34+ (consisting of blood progenitors and stem cells) cell fraction, a step that often improves reprogramming efficiently.  The cell are transfected (nucleofection) with a total of five episome vectors harboring an oriP/EBNA-1 (Epstein-Barr nuclear antigen-1) backbone, individually expressing the reprogramming factors, POU5F1 (OCT4), LIN28A, NANOG, KLF4, CMYC and SOX2. The protocol also includes a phase of transient expression of p53 shRNAs (short inhibitory RNAs) that improve the reprogramming efficiency (Okita et al. 2011). For samples that are difficult to reprogram we boost reprogramming by including an episomal vector carrying an additional expression cassette of CMYC, LIN28A and NANOG (Okita et al. 2011).

The 4in1 method is intended for integration-free reprogramming of human fibroblasts. It is performed via nucleofection of a single vector harbouring polycistronic expression cassette of reprogramming factors, POU5F1 (OCT4), KLF4, SOX2 and CMYC clones in a minicircle plasmid backbone. The protocol also includes a phase of transient expression of p53 shRNAs (short inhibitory RNAs) which improves the reprogramming efficiency (Diecke et al. 2015).