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Gene-disabling techniques simplified by Stanford team

ary to obstruct expression of all genes in a given cell - including genes that were previously uncharacterized. They could then pull individual molecules like books from a shelf to test each one for a biological effect.

The team had several hurdles to overcome in developing their protocol. The first was a size limit - an siRNA molecule longer than 29 subunits causes wide-ranging problems in the cell. The key to overcoming this barrier was a newly available enzyme that snips potential siRNA molecules into 21-subunit lengths. A further step copied these short snippets into a form that could be inserted into a DNA circle called a plasmid. When the researchers put a single plasmid into a cell, it began churning out the gene-blocking siRNA molecule.

The group tested their approach by creating a handful of siRNA molecules to genetically disable three known genes. In each case, their technique generated siRNA that effectively blocked the gene in question. Wehrman said this technique of creating siRNA molecule libraries could be widely used to find genes that, when disabled, cause cells to become cancerous or alter how the cells respond to different drugs. These genes could then become potential targets for drugs to treat disease.

A paper in the same issue of Nature Genetics described a similar way of creating siRNA libraries. "Having two unrelated groups working on the same problem shows there has been a real need for the technology," Blau said. The Stanford group has filed a patent for its technique.


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Contact: Amy Adams
amyadams@stanford.edu
650-723-3900
Stanford University Medical Center
13-Jan-2004


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