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Highly efficient and super-bright neurocircuit tracing using vector mixing-based virus cocktail

By Pei Sun, Sen Jin, Sijue Tao, Junjun Wang, Anan Li, Ning Li, Yang Wu, Jianxia Kuang, Yurong Liu, Lizhao Wang, Huimin Lin, Xiaohua Lv, Xiuli Liu, Jundan Peng, Siyu Zhang, Min Xu, Zhen Luo, Xiaobin He, Tonghui Xu, Xiangning Li, Shaoqun Zeng, Yu-Hui Zhang, Fuqiang Xu

Posted 17 Jul 2019
bioRxiv DOI: 10.1101/705772

Mapping the detailed cell-type-specific input networks and neuronal projectomes are essential to understand brain function in normal and pathological states. However, several properties of current tracing systems, including labeling sensitivity, trans-synaptic efficiencies, reproducibility among different individuals and different Cre-driver animals, still remained unsatisfactory. Here, we developed MAP-ENVIVIDERS, a recombinase system-dependent vector mixing-based strategy for highly efficient neurocircuit tracing. MAP-ENVIVIDERS enhanced tracing efficiency of input networks across the whole brain, with over 10-fold improvement in diverse previously poor-labeled input brain regions and particularly, up to 70-fold enhancement in brainstem compared with the current standard rabies-virus-mediated systems. MAP-ENVIVIDERS was over 10-fold more sensitive for cell-type-specific labeling than previous strategies, enabling us to capture individual cell-type-specific neurons with extremely complex axonal branches and presynaptic axonal boutons, both about one order of magnitude than previously reported and considered. MAP-ENVIVIDERS provides powerful tools for deconstructing novel input/output circuitry towards functional studies and disorders-related mechanisms. ### Competing Interest Statement The authors have declared no competing interest.

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