4.4 Article

Genome editing of pufferfish saxitoxin- and tetrodotoxin-binding protein type 2 in Takifugu rubripes

Journal

TOXICON
Volume 153, Issue -, Pages 58-61

Publisher

PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.toxicon.2018.08.001

Keywords

Takifugu rubripes; Tetrodotoxin; Pufferfish saxitoxin- and tetrodotoxin-binding proteins (PSTBPs); Genome editing; CRISPR/Cas9 system; Knockout

Funding

  1. Japan Society for the Promotion of Science (JSPS) KAKENHI [25660169, 18H02280]

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The pufferfish saxitoxin- and tetrodotoxin-binding protein 2 (PSTBP2), which is involved in toxin accumulation, was knocked out in Takifugu rubripes embryos by using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 genome-editing technology. Treating the embryos with one of two single guide RNA (sgRNA) resulted in mutation rates of 57.1% and 62.5%, respectively, as estimated using a hetero-duplex mobility assay at 3 days postfertilization. Both sgRNAs might induced frameshift mutations that knocked out the T. rubripes PSTBP2.

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