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Single virus used to convert adult cells to embryonic stem cell-like cells
Date:12/15/2008

at gene's protein, stalls when reaching another piece of 2A peptide DNA, and releases the second gene's protein. The process continues until the machinery has made the proteins for all four genes.

Using the tandem genes, Carey created iPS cells containing just a single copy of the polycistronic vector instead of multiple integrations of the viruses. This significant advancement indicates that the approach can become even safer if combined with technologies such as gene targeting, which allows a single transgene to be inserted at defined locations.

Interestingly, while Carey's single-virus method integrates all four genes into the same location, it has proven to be roughly 100 times less efficient than older approaches to reprogramming. This phenomenon remains under investigation.

"We were surprised by the lower efficiency," Carey says. "We're not sure why, but we need to look what's going on with expression levels of the polycistronic virus's proteins compared to separate viruses' proteins."

Although the one virus method is less efficient, Jaenisch maintains it represents an important advance in the field.

"This is an extremely useful tool for studying the mechanisms of reprogramming," says Jaenisch, who is also a professor of biology at MIT. "Using this one virus creates a single integration in the cells' DNA, which makes things much easier to handle."


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Contact: Nicole Giese
giese@wi.mit.edu
617-258-6851
Whitehead Institute for Biomedical Research
Source:Eurekalert

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