TIVA
TIVA
TIVA is a protocol that captures mRNA from live cells. In this method, a photoactivatable TIVA tag is loaded into cells. Selective photoactivation exposes the mRNA-capturing portion of the tag, allowing it to hybridize to the poly(A) tails of mRNA. The biotin-bound mRNA is captured using streptavidin-coated magnetic beads and transcribed into cDNA. Sequencing the cDNA provides transcriptome analysis of RNA from single cells in complex tissues.
Pros:
- In vivo transcriptome analysis using photoactivation in neuronal cells
- Noninvasive method for capturing mRNAs in their natural microenvironment
Cons:
- Limited to a small number of cells
- Zhang X., Marjani S. L., Hu Z., Weissman S. M., Pan X., et al. Single-Cell Sequencing for Precise Cancer Research: Progress and Prospects. Cancer Res. 2016;76:1305-1312
- Binan L., Mazzaferri J., Choquet K., et al. Live single-cell laser tag. Nat Commun. 2016;7:11636
- Bian Q. and Cahan P. Computational Tools for Stem Cell Biology. Trends Biotechnol. 2016;
- Liu S. and Trapnell C. Single-cell transcriptome sequencing: recent advances and remaining challenges. F1000Res. 2016;5:
- Achim K., Pettit J. B., Saraiva L. R., et al. High-throughput spatial mapping of single-cell RNA-seq data to tissue of origin. Nat Biotechnol. 2015;
- Kolodziejczyk A. A., Kim J. K., Svensson V., Marioni J. C. and Teichmann S. A. The Technology and Biology of Single-Cell RNA Sequencing. Mol Cell. 2015;58:610-620
- Baslan T. and Hicks J. Single cell sequencing approaches for complex biological systems. Curr Opin Genet Dev. 2014;26C:59-65