Hosted by the School of Biochemistry

A key requirement for genome engineering is the ability to modularly construct and edit DNA replicons, to precisely programme selective gene expression and to manipulate multiple sets of genes that direct cellular structure and function. To build such programmable genetic circuits, we are developing Large Serine Integrases (LSIs) as orthogonally-acting genome engineering tools that mediate predictable, controllable, and reversible rearrangements of DNA modules. LSIs are enzymes that promote recombination reactions between short distinct sequences of DNA called attachment sites (attB and attP) and they carry out complete DNA cutting and rejoining without leaving any broken ends behind. LSI-catalysed reactions can be seamlessly reversed by the addition of a second protein called a recombination directionality factor (RDF). LSIs have been widely used to promote the insertion of foreign DNA into the genomes of cells, plants and animals. We are using genetic and computational approaches to identify novel RDFs and to study how the LSIs interact with their cognate RDFs. New structural and biochemical insights into LSI-RDF interactions can be applied to develop more flexible specific genome rearrangement events.