Development of synthetic selfish elements based on modular nucleases in Drosophila melanogaster.
Bottom Line: It has been suggested that selfish elements could be exploited to modify the genome of entire populations for medical and ecological applications.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.
Affiliation: Department of Life Sciences, Imperial College London, South Kensington Campus, London SW7 2AZ, UK.Show MeSH
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Mentions: Transgenic fly lines were produced by ϕC31 integrase-mediated insertion into the attP2 docking line (3L: 11 063 638). The TALELAT donor and AAVS1 target were inserted into an otherwise unmarked chromosome. ZFN-AAVS1, ZFN-AAVS1-Long donor and TALELAT target were inserted into an attP2 chromosome marked with curled (cu). The schematic of the homing assay and the possible progeny outcomes from the trans-heterozygous cross depicted in Figure 2B and described in the text in reference to the curled marker exemplifies the situation of the TALELAT line. For ZFN-AAVS1 and ZFN-AAVS1-Long, the curled marker is on the target chromosome, and the progeny was classified accordingly.
Affiliation: Department of Life Sciences, Imperial College London, South Kensington Campus, London SW7 2AZ, UK.