- Xiong, Feng;
- Yang, Hong;
- Song, Yi-Ge;
- Qin, Hai-Bin;
- Zhang, Qing-Yang;
- Huang, Xian;
- Jing, Wei;
- Deng, Manfei;
- Liu, Yang;
- Liu, Zhixiang;
- Shen, Yin;
- Han, Yunyun;
- Lu, Youming;
- Xu, Xiangmin;
- Holmes, Todd C;
- Luo, Minmin;
- Zhao, Fei;
- Luo, Min-Hua;
- Zeng, Wen-Bo
Monosynaptic viral tracers are essential tools for dissecting neuronal connectomes and for targeted delivery of molecular sensors and effectors. Viral toxicity and complex multi-injection protocols are major limiting application barriers. To overcome these barriers, we developed an anterograde monosynaptic H129Amp tracer system based on HSV-1 strain H129. The H129Amp tracer system consists of two components: an H129-dTK-T2-pacFlox helper which assists H129Amp tracer's propagation and transneuronal monosynaptic transmission. The shared viral features of tracer/helper allow for simultaneous single-injection and subsequent high expression efficiency from multiple-copy of expression cassettes in H129Amp tracer. These improvements of H129Amp tracer system shorten experiment duration from 28-day to 5-day for fast-bright monosynaptic tracing. The lack of toxic viral genes in the H129Amp tracer minimizes toxicity in postsynaptic neurons, thus offering the potential for functional anterograde mapping and long-term tracer delivery of genetic payloads. The H129Amp tracer system is a powerful tracing tool for revealing neuronal connectomes.