Wickersham Ian R, Sullivan Heather A
McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139;
Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139.
Cold Spring Harb Protoc. 2015 Apr 1;2015(4):375-85. doi: 10.1101/pdb.prot072389.
Deletion-mutant rabies viral (RV) vectors are powerful tools for neuroscience, allowing monosynaptic tracing of inputs to defined populations and rapid, high-level transgene expression in neurons targeted by multiple routes. High titers and high purity are critical for the successful use of RV vectors in vivo. Here we present a protocol for producing high-quality viral stocks that can be concentrated by ultracentrifugation for final titers in excess of 10(10) infectious units per milliliter.
缺失突变型狂犬病病毒(RV)载体是神经科学研究的有力工具,可用于对特定群体的输入进行单突触追踪,并在多种途径靶向的神经元中实现快速、高水平的转基因表达。高滴度和高纯度对于RV载体在体内的成功应用至关重要。在此,我们介绍一种生产高质量病毒原液的方案,该病毒原液可通过超速离心浓缩,最终滴度超过每毫升10(10)个感染单位。