In:
PLOS ONE, Public Library of Science (PLoS), Vol. 18, No. 3 ( 2023-3-2), p. e0281297-
Abstract:
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 F 0 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.
Type of Medium:
Online Resource
ISSN:
1932-6203
DOI:
10.1371/journal.pone.0281297
DOI:
10.1371/journal.pone.0281297.g001
DOI:
10.1371/journal.pone.0281297.g002
DOI:
10.1371/journal.pone.0281297.g003
DOI:
10.1371/journal.pone.0281297.g004
DOI:
10.1371/journal.pone.0281297.t001
DOI:
10.1371/journal.pone.0281297.t002
DOI:
10.1371/journal.pone.0281297.t003
DOI:
10.1371/journal.pone.0281297.s001
DOI:
10.1371/journal.pone.0281297.s002
DOI:
10.1371/journal.pone.0281297.s003
DOI:
10.1371/journal.pone.0281297.s004
DOI:
10.1371/journal.pone.0281297.s005
DOI:
10.1371/journal.pone.0281297.s006
DOI:
10.1371/journal.pone.0281297.s007
DOI:
10.1371/journal.pone.0281297.s008
DOI:
10.1371/journal.pone.0281297.s009
DOI:
10.1371/journal.pone.0281297.s010
DOI:
10.1371/journal.pone.0281297.s011
DOI:
10.1371/journal.pone.0281297.s012
DOI:
10.1371/journal.pone.0281297.s013
DOI:
10.1371/journal.pone.0281297.r001
DOI:
10.1371/journal.pone.0281297.r002
DOI:
10.1371/journal.pone.0281297.r003
DOI:
10.1371/journal.pone.0281297.r004
Language:
English
Publisher:
Public Library of Science (PLoS)
Publication Date:
2023
detail.hit.zdb_id:
2267670-3
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