Zinc-finger nucleases and targeted insertion of transgenes in cereals

Supervisors: Dr Philip Larkin (02 6246 5060)
Location: Canberra, ACT
Financial support: To be determined

There would be a number of significant advantages achieved in plant biotechnology if transgenes could be targeted to specific sequences in the genome. The benefit of interest for this project is the ability to land transgenes in regions of the chromosome which are already valuable linkage blocks. In this way it would be possible sequentially to add further breeding value to those linkage blocks. New technology is emerging which uses zinc-finger motifs to produce synthetic genes which translate to zinc finger-nucleases which will introduce DNA cuts at sequence-specific locations. In parallel a transgene is introduced with flanking homology to the same site and integration into that site is greatly facilitated by the DNA cut. The project aims to demonstrate the feasibility of this approach to achieve sequence-specific integration in wheat genomes.

The project will involve:

• choosing a target site in alien chromosomal translocations in wheat;
• designing and building zinc-finger nuclease genes for the chosen sequence;
• designing transformation vectors for reporter transgenes; and
• transforming the target cereals and determining the frequency of target and non-target insertions.

In addition similar experiments would be conducted in Arabidopsis and/or rice where the rapidity and ease of transformation would ensure early results for the thesis and serve to guide and confirm the approach for wheat.