A survey of the methodological process of modeling, inference, and evaluation of gene regulatory networks using scRNA-Seq data.

The advent of scRNA-Seq sequencing technology has provided unprecedented resolutions in the analysis of gene regulatory networks (GRNs) at the single-cell level. However, new technical and methodological challenges also emerged. Factors such as the large number of zeros reported in expression levels, the biological variation due to the stochastic nature of gene expression, environmental niche, and effects created by the cell cycle make it difficult to correctly interpret the data obtained in the sequencing stage. On the other hand, the development of methods for the inference of GRNs, specifically using scRNA-Seq technology, proved to be of similar quality to random predictors. The lack of adequate pre-processing of gene expression data, including selection steps for subsets of genes of interest, smoothing, and discretization of gene expression, in addition to the different ways of modeling networks and network motifs, are factors that affect the performance of inference approaches. Finally, the lack of knowledge about the ground-truth network and the non-standardization of appropriate metrics to measure the quality of inferred networks make the process of comparing performance between algorithms a major problem, given the unbalanced nature of the data and the interpretation bias caused by the chosen metric. This article brings these issues to light, aiming to show how these factors influence both the inference process and the performance evaluation of inferred networks, through comparative computational experiments and provides suggestions for a more robust methodological process for researchers dealing with inference of GRNs.