Live Cell Genomics: Cell-Specific Transcriptome Capture in Live Single Cells from Complex Tissues.

Analysis of single-cell transcriptomes shows the single-cell heterogeneity between cells within a population which is vital to our understanding of normal function and disease development. To obtain single-cell transcriptome profiling, however, the poly-A RNA must be accurately isolated from the target cell. We developed a single-cell analysis procedure called transcriptome in vivo analysis (TIVA), which will allow accurate characterization of targeted cell-specific transcriptomes from live tissue. This is accomplished using a RNA capture molecule called TIVA tag that captures the transcriptome of selected cells in their natural microenvironment. An important aspect of the TIVA approach is that the tag is delivered into the cytoplasm of live cells using cell-penetrating peptides (CPPs). Once the TIVA tag is in the cellular cytoplasm, it binds to mRNA after photoactivation of the compound. Using CPPs in combination with photoactivation is the first noninvasive access method for accurately isolating single-cell mRNA from live single cells in tissues in their natural microenvironment.