Transcriptome in vivo analysis (TIVA) of spatially defined single cells in live tissue
Open Access
- 12 January 2014
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature Methods
- Vol. 11 (2), 190-196
- https://doi.org/10.1038/nmeth.2804
Abstract
A noninvasive method is reported for the isolation and profiling of the transcriptome from a single cell in the context of intact tissue. Transcriptome profiling of single cells resident in their natural microenvironment depends upon RNA capture methods that are both noninvasive and spatially precise. We engineered a transcriptome in vivo analysis (TIVA) tag, which upon photoactivation enables mRNA capture from single cells in live tissue. Using the TIVA tag in combination with RNA sequencing (RNA-seq), we analyzed transcriptome variance among single neurons in culture and in mouse and human tissue in vivo. Our data showed that the tissue microenvironment shapes the transcriptomic landscape of individual cells. The TIVA methodology is, to our knowledge, the first noninvasive approach for capturing mRNA from live single cells in their natural microenvironment.Keywords
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