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SV40 DNA could be detected in the offspring but it was impossible to demonstrate with certainty whether the DNA was integrated into the host genome, or remained as episomes .
Bold lines represent yeast chromosomes; circles represent yeast episomes. doi: 10.1371/journal. pone.0010472.g002
The strategy of these authors entailed the introduction of a polycistronic vector encoding the genes c-Myc, Klf4, and SV40LT and selection of stable episomes that provided persistent gene expression to achieve reprogramming followed by the identification of vector-free subclones after several passages in the absence of selection.
"The plasmids carry all the needed transgenes, but don't integrate into the host DNA, they just float around as episomes" in the cell, Thomson says.
"Once the transgenes have done their job and are no longer needed, one can merely recover induced pluripotent stem cells that have lost their episomes."