BATTLE: Genetically Engineered Strategies for Split-Tunable Allocation of Multiple Transgenes in the Nervous System

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. [Display omitted] •Generation of BATTLE-recombinase systems for allocation of multiple transgenes•Split-tunable allocation in BATTLE-1 and multi-sparse allocation in BATTLE-2•Clear and strong labeling of dendrites and axons using BATTLE-2•3D high-resolution imaging of whole synapses in hippocampus in BATTLE-1EX Molecular Biology; Molecular Neuroscience; Neuroanatomy; Techniques in Neuroscience