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Highly efficient transgenic mouse production using piggyBac and its application to rapid phenotyping at the founder generation

Okamura, E.; Mizuno, S.; Matsumoto, S.; Murata, K.; Tanimoto, Y.; Tra, D. T. H.; Suzuki, H.; Kang, W.; Ema, T.; Morimoto, K.; Kato, K.; Matsumoto, T.; Masuyama, N.; Kijima, Y.; Morimura, T.; Sugiyama, F.; Takahashi, S.; Mizutani, E.; Woltjen, K.; Yachie, N.; Ema, M.

2023-12-10 bioengineering
10.1101/2023.12.10.570953 bioRxiv
Show abstract

Pronuclear microinjection is the most popular method for producing transgenic (Tg) animals. Because the production efficiency is typically less than 20%, phenotypic characterization of Tg animals is generally performed on the next generation (F1) onwards. However, apart from in rodents, in many animal species with long generation times, it is desirable to perform phenotyping in the founder (F0) generation. In this study, we attempted to optimize a method of Tg mouse production to achieve higher Tg production efficiency using piggyBac transposon systems and established optimal conditions under which almost all individuals in the F0 generation were Tg. We also succeeded in generating bacterial artificial chromosome Tg mice with efficiency of approximately 70%. By combining this method with genome editing technology, we established a new strategy to perform phenotyping of mice with tissue-specific knockout using the F0 generation. Taking the obtained findings together, by using this method, experimental research using Tg animals can be carried out more efficiently.

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