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Researchers offer proof-of-concept for Altered Nuclear Transfer

dure theorized by Hurlbut is similar to SCNT, but with one crucial twist: Before the donor nucleus is transferred into the egg cell, its DNA is altered so that the resulting blastocyst has no chance of ever becoming a viable embryo. As a result, a "potential human being" is not destroyed once stem cells have been extracted.

Jaenisch--a firm supporter of all forms of human embryonic stem cell research--has shown that technical concerns about this approach can be overcome.

Jaenisch and Alexander Meissner, a graduate student in his lab, focused on a gene called Cdx2, which enables an embryo to grow a placenta. In order to create a blastocyst that cannot implant in a uterus, the researchers disabled Cdx2 in mouse cells.

They accomplished this with a technique called RNA interference, or RNAi. Here, short interfering RNA (siRNA) molecules are designed to target an individual gene and disrupt its ability to produce protein. In effect, the gene is shut off. Jaenisch and Meissner designed a particular form of siRNA that shut off this gene in the donor nucleus and then incorporated itself into all the cells comprising the blastocyst. As a result, all of the resulting mouse blastocysts were incapable of implantation.

However, once the stem cells had been extracted from the blastocysts, Cdx2 was still disabled in each of these new cells, something that needed to be repaired in order for these cells to be useful. To correct this, Meissner deleted the siRNA molecule by transferring a plasmid into each cell. (A plasmid is a unit of DNA that can replicate in a cell apart from the nucleus. Plasmids are usually found in bacteria, and they are a staple for recombinant DNA techniques.) The stem cells resulting from this procedure proved to be just as robust and versatile as stem cells procured in the more traditional fashion.

"The success of this procedure in no way precludes the need to pursue all forms of human embryonic stem cell resea
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Source:Whitehead Institute for Biomedical Research


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