The Advanced capacities of high throughput single cell technologies have facilitated a great understanding of complex biological systems, ranging from cell heterogeneity to molecular expression kinetics. Several pipelines have been introduced to standardize the scRNA-seq analysis workflow. These include cell population identification, cell marker detection and cell trajectory reconstruction. Yet, establishing a systematized pipeline to capture regulatory relationships among transcription factors (TFs) and genes at the cellular level still remains challenging. Here we present PySCNet, a python toolkit that enables reconstructing and analyzing gene regulatory networks (GRNs) from single cell transcriptomic data. PySCNet integrates competitive gene regulatory construction methodologies for cell specific or trajectory specific GRNs and allows for gene co-expression module detection and gene importance evaluation. Moreover, PySCNet offers a user-friendly dashboard website, where GRNs can be customized in an intuitive way.