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One‐step generation of mice with gene editing by Tol2 transposon‐dependent gRNA delivery

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Conditional knockout mice are valuable tools for examining the functions of targeted genes in a time‐ and space‐specific manner. Here, we generated gene‐edited mice by using the Tol2 transposon to… Click to show full abstract

Conditional knockout mice are valuable tools for examining the functions of targeted genes in a time‐ and space‐specific manner. Here, we generated gene‐edited mice by using the Tol2 transposon to introduce guide RNA (gRNA) into fertilized eggs obtained by crossing LSL (loxP‐stop‐loxP)‐CRISPR‐associated 9 (Cas9) mice, which express Cas9 in a Cre‐dependent manner, with CAG‐CreER mice. Transposase mRNA and plasmid DNA, which contained a gRNA sequence for the gene encoding tyrosinase flanked by the transposase recognition sequence, were injected together into fertilized eggs. As a result, the transcribed gRNA cleaved the target genome in a Cas9‐dependent manner. Using this method, it is possible to generate conditional genome‐edited mice more easily in a shorter period of time.

Keywords: tol2 transposon; grna; one step; step generation; gene

Journal Title: FEBS Letters
Year Published: 2023

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