A transcript is an RNA molecule that is copied or transcribed from a DNA template. A transcript can be further processed by alternative splicing, which is the retention of different combinations of exons. These unique combinations of exons are termed RNA transcript isoforms. The transcriptome is a set of all RNA, including rRNA, mRNA, tRNA, and non-coding RNA. Specifically mRNA transcripts can be used to investigate differences in gene expression patterns. Transcriptome profiling is determining the composition of transcripts and their relative expression levels in a given reference set of cells. This analysis involves characterization of all functional genomic elements, coding and non-coding.
The current RNA capture methods involve sorting cells in suspension from acutely dissociated tissue, and thus can lose information about cell morphology and microenvironment. Transcript abundance and isoforms are significantly different across tissues and are continually changing throughout an individual’s life. Gene expression is highly tissue specific, therefore with traditional RNA capture methods one must be cautious in the interpretation of gene expression patterns, as they often reflect expression of a heterogeneous mix of cell populations. Even in the same cell type, tissue measurements, where a population of cells is obtained, mask both low-level mRNA expression in single cells and variation in expression between cells. The photoactivatable TIVA tag is engineered to capture the mRNA of a single cell in complex tissues.