Split selectable marker systems utilizing inteins facilitate gene stacking in plants
Scientists from the Oak Ridge National Laboratory (ORNL) successfully inserted multiple genes into plants using only a single step. The new method could revolutionize the way researchers around the world conduct genetic transformation.
The technique, known as gene stacking, replaces the laborious way of individually inserting genes into a targeted plant's DNA. Each transformation also requires its own confirmation test to determine if the gene is in the right spot and orientation to exhibit the intended desired trait. Gene stacking allows scientists to do multiple gene insertions and confirmation tests in just one transformation.
The new delivery method uses intein protein segments, which can naturally split off from larger proteins then splice back together to create new proteins. The inteins were used to create a split selectable marker system that simultaneously inserted four genes, including markers, into plants. Gene stacking was successfully demonstrated and confirmed using tobacco, Arabidopsis thaliana, and poplar.
ORNL scientists are already modifying the technique to allow 12 gene insertions at once, two of which are marker genes. They are optimistic that their gene stacking method can support as many as 20 gene insertions at a single transformation.