Figure 1
From: TALEN-mediated knock-in via non-homologous end joining in the crustacean Daphnia magna
Donor plasmid design for knock-in into the eyeless locus.
(a) Exon-intron structure of the eyeless (ey) gene in D. magna. A predicted structure of ey is schematically represented based on sequence data obtained from D. magna public gene set data and database (http://arthropods.eugenes.org/EvidentialGene/daphnia/daphnia_magna_new/). Exons and introns are shown in black and white respectively, with the exception of ey1 TALEN targeting exon 7 (red) and ey2 TALEN targeting exon 9 (blue). The coding sequence from exon 7 to exon 10 was confirmed experimentally in our previous study24. Arrowheads indicate ey1 TALEN and ey2 TALEN cleavage sites. Genomic regions for the insertion of donor DNA by ey1 TALEN target and ey2 TALEN target are shown in brackets. (b) Schematic representation of donor DNA. The H2B-GFP ORF attached to D. magna EF1a1 3′UTR was used as a reporter. Four tandem repeats of the JHRE element were inserted upstream of the reporter, which was also flanked with the ey1 (red) and ey2 (blue) TALEN target sequences.
