Transcriptome In Vivo Analysis (TIVA) of spatially defined single cells in intact live mouse and human brain tissue

Details

• Alternative Title:
  Nat Methods
• Personal Author:
  Lovatt, Ditte ; Ruble, Brittani K. ; Lee, Jaehee ; Dueck, Hannah ; Kim, Tae Kyung ; Fisher, Stephen ; Francis, Chantal ; Spaethling, Jennifer M. ; Wolf, John A. ; Grady, M. Sean ; Ulyanova, Alexandra V. ; Yeldell, Sean B. ; Griepenburg, Julianne C. ; Buckley, Peter T. ; Kim, Junhyong ; Sul, Jai-Yoon ; Dmochowski, Ivan J. ; Eberwine, James
• Description:
  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:
  Animals Brain Computational Biology Gene Expression Profiling Gene Library High-Throughput Nucleotide Sequencing Hippocampus Humans Mice Mice, Inbred C57BL Neurons RNA, Messenger Sequence Analysis, RNA

  More +
• Source:
  Nat Methods. 11(2):190-196
• Pubmed ID:
  24412976
• Pubmed Central ID:
  PMC3964595
• Document Type:
  Journal Article
• Funding:
  R01 GM083030/GM/NIGMS NIH HHSUnited States/ ; S10 RR021113/RR/NCRR NIH HHSUnited States/ ; U01 MH098953/MH/NIMH NIH HHSUnited States/ ; DP004117/DP/NCCDPHP CDC HHSUnited States/ ; DP1 OD004117/OD/NIH HHSUnited States/ ; DP1 MH099902/MH/NIMH NIH HHSUnited States/ ; U01MH098953/MH/NIMH NIH HHSUnited States/ ; U58 DP004117/DP/NCCDPHP CDC HHSUnited States/
• Volume:
  11
• Issue:
  2
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:5f6e9dae0c29c4490c351523f935c1aa075496e786cb775515efe6cfce1aec2f
• Download URL:
  https://stacks.cdc.gov/view/cdc/24457/cdc_24457_DS1.pdf
• File Type:
  [PDF - 1.22 MB ]