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. 2020 Aug 31;15(8):e0238256. doi: 10.1371/journal.pone.0238256

Fig 1. Genotype of the two Dsx1-related mutants.

Fig 1

A) Structure of Dsx1 gene of Daphnia magna and the chosen design of TALEN for genome editing. There are two different promoters, alpha and beta, directing the transcription of two Dsx1 transcripts which are different only in 5’ UTR region. TALEN was designed to target a 46 bp sequence at the beginning of the common exon 4 which contains the full ORF of Dsx1. Cleavage is expected to occur right in front of Dsx1 start codon. A donor vector carrying a copy of promoter-less mCherry was also used in this targeting strategy. The 46-bp TALEN target sequence is cloned into 5’ end and the full-length 3’ UTR of Dsx1 is cloned into 3’ end of this mCherry to allow NHEJ-mediated knock-in so that mCherry can replace Dsx1 ORF and report the activity of the locus. B) Diagram illustrating the genotype at Dsx1 locus of the two mutant strains, Line A and Line B. Both are hemizygous knock-ins with in-del mutations found at TALEN cleavage site. C) Nucleotide sequence of in-del mutations from the four alleles shown in panel B). At the bottom, the full amino acid sequence of Dsx1 protein is shown together with the position of in-frame methionines that may become the alternative start codon in case the original one is deleted.