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Transcriptome in vivo analysis (TIVA) of spatially defined single cells in live tissue.
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.
Afiliação
  • Lovatt D; 1] Department of Pharmacology, University of Pennsylvania, Philadelphia, Pennsylvania, USA. [2].
  • Ruble BK; 1] Department of Chemistry, University of Pennsylvania, Philadelphia, Pennsylvania, USA. [2].
  • Lee J; Department of Pharmacology, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
  • Dueck H; Department of Biology, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
  • Kim TK; Department of Pharmacology, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
  • Fisher S; Department of Biology, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
  • Francis C; Department of Biology, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
  • Spaethling JM; Department of Pharmacology, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
  • Wolf JA; Department of Neurosurgery, University of Pennsylvania Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
  • Grady MS; Department of Neurosurgery, University of Pennsylvania Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
  • Ulyanova AV; Department of Neurosurgery, University of Pennsylvania Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
  • Yeldell SB; Department of Chemistry, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
  • Griepenburg JC; Department of Chemistry, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
  • Buckley PT; Department of Pharmacology, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
  • Kim J; 1] Department of Biology, University of Pennsylvania, Philadelphia, Pennsylvania, USA. [2] PENN Genome Frontiers Institute, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
  • Sul JY; Department of Pharmacology, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
  • Dmochowski IJ; 1] Department of Chemistry, University of Pennsylvania, Philadelphia, Pennsylvania, USA. [2].
  • Eberwine J; 1] Department of Pharmacology, University of Pennsylvania, Philadelphia, Pennsylvania, USA. [2] PENN Genome Frontiers Institute, University of Pennsylvania, Philadelphia, Pennsylvania, USA. [3].
Nat Methods ; 11(2): 190-6, 2014 Feb.
Article em En | MEDLINE | ID: mdl-24412976
ABSTRACT
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.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Encéfalo / Análise de Sequência de RNA / Perfilação da Expressão Gênica / Sequenciamento de Nucleotídeos em Larga Escala / Hipocampo / Neurônios Limite: Animals / Humans Idioma: En Revista: Nat Methods Assunto da revista: TECNICAS E PROCEDIMENTOS DE LABORATORIO Ano de publicação: 2014 Tipo de documento: Article
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Encéfalo / Análise de Sequência de RNA / Perfilação da Expressão Gênica / Sequenciamento de Nucleotídeos em Larga Escala / Hipocampo / Neurônios Limite: Animals / Humans Idioma: En Revista: Nat Methods Assunto da revista: TECNICAS E PROCEDIMENTOS DE LABORATORIO Ano de publicação: 2014 Tipo de documento: Article