RECOMBINASES FOR INTEGRATING DNA & RECOMBINASE FUSIONS

Manipulation of eukaryotic genomes, particularly the integration of multi-kilobase DNA
sequences, remains challenging and limits the rapidly growing fields of synthetic biology and
cell engineering. Large serine recombinases (LSRs) are enzymes that recognize specific target
sequences on a DNA donor sequence and DNA target sequence to catalyze a recombination
reaction that results in the insertion of a DNA donor in a sequence-specific manner. Genome
editing can be carried out using an LSR system and a DNA donor nucleic acid, such as a plasmid
or double-stranded DNA. However, in a human genome, these systems can exhibit variable
efficiency and specificity.

In this invention, UC Berkeley researchers and others have developed optimized compositions
to significantly increase the efficiency and specificity of LSRs to target a specific genomic locus
in human cells. Via fusion to additional protein systems, this engineered composition retains
the simplicity of a single protein for gene delivery. The invention also encompasses use in in
vivo or ex vivo gene therapy and the creation of modified cell lines or transgenic animals.