Median fins are thought to be ancestors of paired fins which in turn give rise to limbs in tetrapods. However, the developmental mechanisms of median fins remain largely unknown. Nonsense mutation of the T-box transcription factor eomesa in zebrafish results in a phenotype without dorsal fin. Compared to zebrafish, the common carp undergo an additional round of whole genome duplication, acquiring an extra copy of protein-coding genes. To verify the function of eomesa genes in common carp, we established a biallelic gene editing technology in this tetraploidy fish through simultaneous disruption of two homologous genes, eomesa1 and eomesa2. We targeted four sites located upstream or within the sequences encoding the T-box domain. Sanger sequencing data indicated the average knockout efficiency was around 40% at T1-T3 sites and 10% at T4 site in embryos at 24 hours post fertilization. The individual editing efficiency was high to about 80% at T1-T3 sites and low to 13.3% at T4 site in larvae at 7 days post fertilization. Among 145 mosaic F0 examined at four months old, three individuals (Mutant 1-3) showed varying degrees of maldevelopment in the dorsal fin and loss of anal fin. Genotyping showed the genomes of all three mutants were disrupted at T3 sites. The null mutation rates on the eomesa1 and eomesa2 loci were 0% and 60% in Mutant 1, 66.7% and 100% in Mutant 2, and 90% and 77.8% in Mutant 3, respectively. In conclusion, we demonstrated a role of eomesa in the formation and development of median fins in Oujiang color common carp and established an method that simultaneously disrupt two homologous genes with one gRNA, which would be useful in genome editing in other polyploidy fishes.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0281297 | PLOS |
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Institute of Marine Biology, National Taiwan Ocean University, Keelung 202, Taiwan National Taiwan Ocean University Keelung Taiwan.
Two new species of dark-body snake eels are described based on specimens collected from Taiwan. has a long tail; dorsal-fin origin above posterior third of pectoral fin; tip of lower jaw anterior to anterior-nostril tube; two simple, pointed protrusions along upper lip; preoperculomandibular pores 6 or 7 + 3; teeth on jaws and vomer mostly uniserial, except for biserial on posterior portion of maxilla and anterior portion of symphysis of dentary; vertebral formula 12-55-153 and median fins with narrow dark margins, except the pale fin origins. has a dorsal-fin origin well behind gill opening; mainly 4 rows of teeth on jaws; no protrusions along upper lip; a smaller head; mean vertebral formula 24-64-163 and pale median fins.
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Department of Craniofacial Biology, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA.
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Department of Ecological Developmental Adaptability Life Sciences, Graduate School of Life Sciences, Tohoku University, Sendai 980-8578, Japan.
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