Following single-cell isolation, the epigenome and the transcriptome may also be studied [21]. While epigenomics of single cells remains challenging [52, 53, 54 and 55], methods for single-cell transcriptomics have flourished (Figure 4) and delivered baffling new insight into the (functional) heterogeneity of cell populations [56, 57 and 58]. A single cell contains less than 1 pg of mRNA. To characterize it via array [59 and 60] or sequencing [56, 57 and 58] approaches, whole-transcriptome amplification (WTA) is required. Methods for WTA are grounded on PCR-based [61, 62, 63, 64, 65•, 66, 67, 68 and 69], MDA-based

[67] or in vitro transcription (IVT)-based [ 70] amplification LBH589 purchase of reverse transcribed single-cell mRNA, whereby IVT likely results in a more linear amplification. However, WTA and subsequent analysis methods struggle with reliable amplification and detection of transcripts expressed at less than 10 copies per cell. In addition, the majority of the methods only selectively amplify the polyadenylated RNAs of a cell’s transcript repertoire [ 61, 62, 63, 64, 65•, 66 and 70], and may be biased to the 3′-end [ 70] or the 5′-end [ 61 and 62]

of a transcript ( Figure 4). Full-length mRNA-characterization from a IBET762 single cell can only be achieved by a few WTA-methods [ 63, 64, 65•, 66, 67 and 68] ( Figure 4). To the best of our knowledge, no large-scale single cancer cell transcriptome sequencing studies have been reported, although

this is on the horizon [71, 72 and 73]. In a recent elegant proof-of-principle experiment, Ramsköld et al. found differences between melanoma CTCs and primary melanocytes, giving insight into the disease [ 65•]. Additionally, the technology allowed defining potent plasma membrane CTC biomarkers and discovering expressed coding mutations. This and other studies [ 60, 73, 74 and 75] show that single-cell transcriptomics will illuminate further insights into oncogenesis, tumour subclonal architecture and cell lineage diversity. Single-cell sequencing studies currently only process either WGA-products or WTA-products of a cell, although protocols for combined approaches are under development [76• and 77]. The ability to profile both the genome and the transcriptome of the same cell has enormous potential to elucidate Astemizole heterogeneity at the genome, epigenome and transcriptome level. In addition, such techniques would allow mutations of the genome in a single cell to be confirmed in that cell’s transcriptome, opening avenues to detect mutations at high confidence, even if they are observed only in one single cell. The emerging field of single cell genomics opens new avenues that may have far-reaching implications for cancer research and clinical practice. It allows characterization of intra-tumour genetic heterogeneity genome-wide to single-cell resolution, and thereby offers a unique viewpoint into tumour evolution.