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


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Abstract

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 heteroduplex mobility assay at 3 days postfertilization. Both sgRNAs might induced frameshift mutations that knocked out the T. rubripes PSTBP2.HIGHLIGHTSThe CRISPR/Cas9 system was used to knock out the PSTBP2 gene in Takifugu rubripes.Off-target effects occurred at very low levels, or were not detected.This knockout is useful for studying the mechanism of toxin accumulation in fishes.

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