From silencing to gene expression: Real-time analysis in single cells

Susan M. Janicki, Toshiro Tsukamoto, Simone E. Salghetti, William P. Tansey, Ravi Sachidanandam, Kannanganattu V. Prasanth, Thomas Ried, Yaron Shav-Tal, Edouard Bertrand, Robert H. Singer, David L. Spector

Research output: Contribution to journalArticle

473 Citations (Scopus)

Abstract

We have developed an inducible system to visualize gene expression at the levels of DNA, RNA and protein in living cells. The system is composed of a 200 copy transgene array integrated into a euchromatic region of chromosome 1 in human U2OS cells. The condensed array is heterochromatic as it is associated with HP1, histone H3 methylated at lysine 9, and several histone methyltransferases. Upon transcriptional induction, HP1α is depleted from the locus and the histone variant H3.3 is deposited suggesting that histone exchange is a mechanism through which heterochromatin is transformed into a transcriptionally active state. RNA levels at the transcription site increase immediately after the induction of transcription and the rate of synthesis slows over time. Using this system, we are able to correlate changes in chromatin structure with the progression of transcriptional activation allowing us to obtain a real-time integrative view of gene expression.

Original languageEnglish (US)
Pages (from-to)683-698
Number of pages16
JournalCell
Volume116
Issue number5
DOIs
StatePublished - Mar 5 2004

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Single-Cell Analysis
Gene expression
Histones
Transcription
Gene Expression
RNA
Heterochromatin
Chromosomes, Human, Pair 1
Chromosomes
Transgenes
Transcriptional Activation
Lysine
Chromatin
Chemical activation
Cells
DNA
Proteins

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology

Cite this

Janicki, S. M., Tsukamoto, T., Salghetti, S. E., Tansey, W. P., Sachidanandam, R., Prasanth, K. V., ... Spector, D. L. (2004). From silencing to gene expression: Real-time analysis in single cells. Cell, 116(5), 683-698. https://doi.org/10.1016/S0092-8674(04)00171-0

From silencing to gene expression : Real-time analysis in single cells. / Janicki, Susan M.; Tsukamoto, Toshiro; Salghetti, Simone E.; Tansey, William P.; Sachidanandam, Ravi; Prasanth, Kannanganattu V.; Ried, Thomas; Shav-Tal, Yaron; Bertrand, Edouard; Singer, Robert H.; Spector, David L.

In: Cell, Vol. 116, No. 5, 05.03.2004, p. 683-698.

Research output: Contribution to journalArticle

Janicki, SM, Tsukamoto, T, Salghetti, SE, Tansey, WP, Sachidanandam, R, Prasanth, KV, Ried, T, Shav-Tal, Y, Bertrand, E, Singer, RH & Spector, DL 2004, 'From silencing to gene expression: Real-time analysis in single cells', Cell, vol. 116, no. 5, pp. 683-698. https://doi.org/10.1016/S0092-8674(04)00171-0
Janicki SM, Tsukamoto T, Salghetti SE, Tansey WP, Sachidanandam R, Prasanth KV et al. From silencing to gene expression: Real-time analysis in single cells. Cell. 2004 Mar 5;116(5):683-698. https://doi.org/10.1016/S0092-8674(04)00171-0
Janicki, Susan M. ; Tsukamoto, Toshiro ; Salghetti, Simone E. ; Tansey, William P. ; Sachidanandam, Ravi ; Prasanth, Kannanganattu V. ; Ried, Thomas ; Shav-Tal, Yaron ; Bertrand, Edouard ; Singer, Robert H. ; Spector, David L. / From silencing to gene expression : Real-time analysis in single cells. In: Cell. 2004 ; Vol. 116, No. 5. pp. 683-698.
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