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Scientists report first transgenic animal developed via retroviral DNA insertion into male germ-line stem cells

life.

Of the various types of stem cells, only two spermatogonial and hematopoeitic stem cells can be positively identified using functional assays. This makes them valuable models for other types of stem cells, such as those that give rise to skin, the lining of the intestines, brain, muscle and liver.

Spermatogonial stem cells are of additional interest for transgenics applications because they are the only cells, including all other stem cells, that undergo self-renewal throughout an animals lifetime and contribute genes to subsequent generations. Previous attempts to genetically alter this unique type of stem cell, either through retroviruses or other methods, have met with little success.

Brinsters work is unusual in that his transgenic mice were created using cells from male animals. Transgenic animals are generally produced by inserting foreign DNA into cells derived from females, such as oocytes, eggs and blastocysts. Brinsters group has demonstrated that the efficiency of generating transgenic mice through spermatogonial stem cells is roughly equivalent to that of female-derived cells.

"Questions had been raised regarding whether male germ-line stem cells could be transduced with a retroviral vector and whether any gene introduced would be silenced," Brinster said. "Our work clearly demonstrates that the stem cell can be transduced at relatively high efficiency and expression is not silenced. About 10 percent of stem cells carry active genes that are transmitted for at least three generations."

Retroviruses are the most common vehicles for introducing genes in human somatic cell gene therapy, and some scientists had expressed concern that this approach might result in genetic alterations to germ-line cells. Brinsters paper indicates that the germ cells are indeed susceptible to insertion of foreign genes via retroviruses, although the somatic cells that surround stem cells in the body most li
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Contact: Steve Bradt
bradt@pobox.upenn.edu
215-573-6604
University of Pennsylvania
22-Oct-2001


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