title: Development of synthetic selfish elements based on modular nucleases in Drosophila melanogaster.
abstract: Selfish genes are DNA elements that increase their rate of genetic transmission at the expense of other genes in the genome and can therefore quickly spread within a population. It has been suggested that selfish elements could be exploited to modify the genome of entire populations for medical and ecological applications. Here we report that transcription activator-like effector nuclease (TALEN) and zinc finger nuclease (ZFN) can be engineered into site-specific synthetic selfish elements (SSEs) and demonstrate their transmission of up to 70% in the Drosophila germline. We show here that SSEs can spread via DNA break-induced homologous recombination, a process known as 'homing' similar to that observed for homing endonuclease genes (HEGs), despite their fundamentally different modes of DNA binding and cleavage. We observed that TALEN and ZFN have a reduced capability of secondary homing compared to HEG as their repetitive structure had a negative effect on their genetic stability. The modular architecture of ZFNs and TALENs allows for the rapid design of novel SSEs against specific genomic sequences making them potentially suitable for the genetic engineering of wild-type populations of animals and plants, in applications such as gene replacement or population suppression of pest species.
authors.affiliation: Department of Life Sciences, Imperial College London, South Kensington Campus, London SW7 2AZ, UK.
authors.forename: Alekos
authors.initials: A
authors.lastname: Simoni
authors.affiliation: Department of Life Sciences, Imperial College London, South Kensington Campus, London SW7 2AZ, UK.
authors.forename: Carla
authors.initials: C
authors.lastname: Siniscalchi
authors.affiliation: Department of Genetics, University of Cambridge, Downing Street, Cambridge CB2 3EH, UK.
authors.forename: Yuk-Sang
authors.initials: YS
authors.lastname: Chan
authors.affiliation: Department of Genetics, University of Cambridge, Downing Street, Cambridge CB2 3EH, UK.
authors.forename: David S
authors.initials: DS
authors.lastname: Huen
authors.affiliation: Department of Genetics, University of Cambridge, Downing Street, Cambridge CB2 3EH, UK.
authors.forename: Steven
authors.initials: S
authors.lastname: Russell
authors.affiliation: Department of Life Sciences, Imperial College London, South Kensington Campus, London SW7 2AZ, UK.
authors.forename: Nikolai
authors.initials: N
authors.lastname: Windbichler
authors.affiliation: Department of Life Sciences, Imperial College London, South Kensington Campus, London SW7 2AZ, UK Centre of FunctionalGenomics, University of Perugia, S. Andrea delle Fratte, 06132, Perugia, Italy a.crisanti@imperial.ac.uk.
authors.forename: Andrea
authors.initials: A
authors.lastname: Crisanti
mesh_terms: D000818:Animals; D004247:DNA / Q000737:chemistry; D059766:DNA End-Joining Repair; D004331:Drosophila melanogaster / Q000235:genetics*; D004706:Endodeoxyribonucleases / Q000235:genetics* / Q000378:metabolism; D005260:Female; D059765:Homologous Recombination; D008297:Male; D015202:Protein Engineering
chemical_list: D004247:DNA; D004706:Endodeoxyribonucleases