HSV-1 H129-Derived Anterograde Neural Circuit Tracers: Improvements, Production, and Applications

Hong Yang 1,2 • Feng Xiong 1,2 • Yi-Ge Song 1,2 • Hai-Fei Jiang 1,2 • Hai-Bin Qin 1,2 • Jing Zhou 1,2 • Sha Lu 3 • Steven F. Grieco 4 • Xiangmin Xu 4 • Wen-Bo Zeng 1 • Fei Zhao 5,6 • Min-Hua Luo 1,2,7

1 State Key Laboratory of Virology, CAS Center for Excellence in Brain Science and Intelligence Technology, Center for Biosafety Mega-Science, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China 

2 University of Chinese Academy of Sciences, Beijing 100049, China 

3 Shanghai Genechem Co. Ltd., Shanghai 201203, China 

4 Department of Anatomy and Neurobiology, School of Medicine, University of California, Irvine, CA 92697, USA 

5 School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China 

6 Chinese Institute for Brain Research, Beijing 102206, China 

7 Shanghai Public Health Clinical Center, Fudan University, Shanghai 200032, China




Anterograde viral tracers are powerful and essential tools for dissecting the output targets of a brain region of interest. They have been developed from herpes simplex virus 1 (HSV-1) strain H129 (H129), and have been successfully applied to map diverse neural circuits. Initially, the anterograde polysynaptic tracer H129-G4 was used by many groups. We then developed the first monosynaptic tracer, H129-dTK-tdT, which was highly successful, yet improvements are needed. Now, by inserting another tdTomato expression cassette into the H129-dTK-tdT genome, we have created H129-dTK-T2, an updated version of H129-dTK-tdT that has improved labeling intensity. To help scientists produce and apply our H129-derived viral tracers, here we provide the protocol describing our detailed and standardized procedures. Commonly-encountered technical problems and their solutions are also discussed in detail. Broadly, the dissemination of this protocol will greatly support scientists to apply these viral tracers on a large scale.



HSV-1 strain H129 (H129) ; Anterograde transneuronal tracer ; Polysynaptic tracer ; Monosynaptic tracer; Production ; Application; Neural circuit ; Neural circuit tracing