Single cell studies increasing reveal myriad cellular subtypes beyond those postulated or observed through optical and fluorescence microscopy as well as DNA sequencing studies. While gene sequencing at the single cell level offer a path towards illuminating, in totality, the different subtypes of cells present, the technique nevertheless does not offer answers concerning the functional repertoire of the cell, which is defined by the collection of RNA transcribed from the genome. Known as the transcriptome, transcribed RNA defines the function of the cell as proteins or effector RNA molecules, while the genome is the collection of all information endowed in the cell type, expressed or not. Thus, a particular cell state, lineage, cell fate or cellular differentiation is more fully depicted by transcriptomic analysis compared to delineating the genomic context at the single cell level. While conceptually sound and could be analysed by contemporary single cell RNA sequencing technology and data analysis pipelines, the relative instability of RNA in view of RNase in the environment would make sample preparation particularly challenging, where degradation of cellular RNA by extraneous factors could provide a misinterpretation of specific functions available to a cell type. Hence, RNA as the de facto functional molecule of the cell defining the proteomics landscape as well as effector RNA repertoire, meant that RNA transcriptomics at the single cell level is the way forward if the goal is to understand all available cell types, lineage, cell fate and cellular differentiation. Given that a cell state is defined by the functions encoded by functional molecules such as proteins and RNA, single cell RNA sequencing offers a larger contextual basis for understanding cellular decision making and functions, for example, proteins are increasingly known to work in concert with RNA effector molecules in enabling a function. Hence, providing a view of the diverse cell types and lineages present in a body, single cell RNA sequencing is only hampered by the high sensitivity required to analyse the small amount of RNA available in single cells, as well as the perennial problem of RNA studies: how to prevent or reduce RNA degradation by environmental RNase enzymes. Ability to reduce RNA degradation would provide the cell biologist a unique view of the functional landscape of different cells in the body through the language of RNA.
Single-Cell Analysis for Whole-Organism Datasets
