Gene Targeting Without Nucleases

Researchers in Dr. Mark Kay's lab have developed a method for site-specific gene targeting that does not require nucleases. Gene therapy attempts to cure disease by manipulating the genetic material which holds the instructions for the function of our cells and our bodies. A major challenge in gene therapy is to deliver therapeutic genes to desired places in the genome with as little possible off target effects. Previous efforts to manipulate the genome have relied on the use of molecular scissors called nucleases to increase the rate of site-specific integration. However, these targeted nucleases are often toxic to cells and their off target activity may be immunogenic and genotoxic. To help overcome these challenges, the inventors have developed this method. The method does not use nucleases, a vector-borne promoter or disrupt the expression of the genomic gene at the integration site and thus provides a superior safety profile.

Stage of research
The investigators have used this method to correct the genetic disease Hemophilia B in mice. In addition to curing the mice, the inventors showed that the therapeutic gene was expressed specifically from the desired place and that the expression of the genomic gene at the integration site was not disrupted.