BATTLE: Genetically Engineered Strategies for Split-Tunable Allocation of Multiple Transgenes in the Nervous System
Keigo Kohara,
Akitoshi Inoue,
Yousuke Nakano,
Hirokazu Hirai,
Takuya Kobayashi,
Masato Maruyama,
Ryosuke Baba,
Chiho Kawashima
Affiliations
Keigo Kohara
Department of Cellular and Functional Biology, Institute of Biomedical Science, Kansai Medical University, Hirakata, Osaka 573-1010, Japan; Corresponding author
Akitoshi Inoue
Department of Medical Chemistry, Kansai Medical University, Graduate School of Medicine, Hirakata, Osaka 573-1010, Japan
Yousuke Nakano
Department of Anatomy, Kansai Medical University, Graduate School of Medicine, Hirakata, Osaka 573-1010, Japan
Hirokazu Hirai
Department of Neurophysiology and Neural Repair, Gunma University Graduate School of Medicine, Maebashi, Gunma 371-8511, Japan; Research Program for Neural Signalling, Division of Endocrinology, Metabolism and Signal Research, Gunma University Initiative for Advanced Research, Maebashi, Gunma 371-8512, Japan
Takuya Kobayashi
Department of Medical Chemistry, Kansai Medical University, Graduate School of Medicine, Hirakata, Osaka 573-1010, Japan; Japan Agency for Medical Research and Development (AMED), Core Research for Evolutional Science and Technology (CREST), 2-5-1 Shinmachi, Hirakata, Osaka 573-1010, Japan
Masato Maruyama
Department of Anatomy, Kansai Medical University, Graduate School of Medicine, Hirakata, Osaka 573-1010, Japan; Faculty of Pharmaceutical Sciences, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama 700-8530, Japan
Ryosuke Baba
Department of Cellular and Functional Biology, Institute of Biomedical Science, Kansai Medical University, Hirakata, Osaka 573-1010, Japan
Chiho Kawashima
Department of Cellular and Functional Biology, Institute of Biomedical Science, Kansai Medical University, Hirakata, Osaka 573-1010, Japan; Department of Bioscience, Osaka College of High Technology, Osaka 532-003, Japan
Summary: Elucidating fine architectures and functions of cellular and synaptic connections requires development of new flexible methods. Here, we created a concept called the “battle of transgenes,” based on which we generated strategies using genetically engineered battles of multiple recombinases. The strategies enabled split-tunable allocation of multiple transgenes. We demonstrated the versatility of these strategies and technologies in inducing strong and multi-sparse allocations of multiple transgenes. Furthermore, the combination of our transgenic strategy and expansion microscopy enabled three-dimensional high-resolution imaging of whole synaptic structures in the hippocampus with simultaneous visualizations of endogenous synaptic proteins. These strategies and technologies based on the battle of genes may accelerate the analysis of whole synaptic and cellular connections in diverse life science fields.
