Use of the fluorescent timer DsRED-E5 as reporter to monitor dynamics of gene activity in plants

R. Mirabella, C. Franken, G.N.M. van der Krogt, T. Bisseling, R. Geurts

Research output: Contribution to journalArticleAcademicpeer-review

36 Citations (Scopus)

Abstract

Fluorescent proteins, such as green fluorescent protein and red fluorescent protein (DsRED), have become frequently used reporters in plant biology. However, their potential to monitor dynamic gene regulation is limited by their high stability. The recently made DsRED-E5 variant overcame this problem. DsRED-E5 changes its emission spectrum over time from green to red in a concentration independent manner. Therefore, the green to red fluorescence ratio indicates the age of the protein and can be used as a fluorescent timer to monitor dynamics of gene expression. Here, we analyzed the potential of DsRED-E5 as reporter in plant cells. We showed that in cowpea (Vigna unguiculata) mesophyll protoplasts, DsRED-E5 changes its fluorescence in a way similar to animal cells. Moreover, the timing of this shift is suitable to study developmental processes in plants. To test whether DsRed-E5 can be used to monitor gene regulation in plant organs, we placed DsRED-E5 under the control of promoters that are either up- or down-regulated (MtACT4 and LeEXT1. promoters) or constitutively expressed (MtACT2 promoter) during root hair development in Medicago truncatula. Analysis of the fluorescence ratios clearly provided more accurate insight into the timing of promoter activity.
Original languageEnglish
Pages (from-to)1879-1887
JournalPlant Physiology
Volume135
Issue number4
DOIs
Publication statusPublished - 2004

Fingerprint

timers
Fluorescence
promoter regions
monitoring
fluorescence
Medicago truncatula
Genes
genes
Protoplasts
Plant Cells
Green Fluorescent Proteins
plant biology
Proteins
root hairs
Vigna unguiculata
plant organs
green fluorescent protein
cowpeas
mesophyll
protoplasts

Keywords

  • medicago-truncatula
  • protein gfp
  • expression
  • arabidopsis
  • marker
  • rna

Cite this

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title = "Use of the fluorescent timer DsRED-E5 as reporter to monitor dynamics of gene activity in plants",
abstract = "Fluorescent proteins, such as green fluorescent protein and red fluorescent protein (DsRED), have become frequently used reporters in plant biology. However, their potential to monitor dynamic gene regulation is limited by their high stability. The recently made DsRED-E5 variant overcame this problem. DsRED-E5 changes its emission spectrum over time from green to red in a concentration independent manner. Therefore, the green to red fluorescence ratio indicates the age of the protein and can be used as a fluorescent timer to monitor dynamics of gene expression. Here, we analyzed the potential of DsRED-E5 as reporter in plant cells. We showed that in cowpea (Vigna unguiculata) mesophyll protoplasts, DsRED-E5 changes its fluorescence in a way similar to animal cells. Moreover, the timing of this shift is suitable to study developmental processes in plants. To test whether DsRed-E5 can be used to monitor gene regulation in plant organs, we placed DsRED-E5 under the control of promoters that are either up- or down-regulated (MtACT4 and LeEXT1. promoters) or constitutively expressed (MtACT2 promoter) during root hair development in Medicago truncatula. Analysis of the fluorescence ratios clearly provided more accurate insight into the timing of promoter activity.",
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author = "R. Mirabella and C. Franken and {van der Krogt}, G.N.M. and T. Bisseling and R. Geurts",
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Use of the fluorescent timer DsRED-E5 as reporter to monitor dynamics of gene activity in plants. / Mirabella, R.; Franken, C.; van der Krogt, G.N.M.; Bisseling, T.; Geurts, R.

In: Plant Physiology, Vol. 135, No. 4, 2004, p. 1879-1887.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Use of the fluorescent timer DsRED-E5 as reporter to monitor dynamics of gene activity in plants

AU - Mirabella, R.

AU - Franken, C.

AU - van der Krogt, G.N.M.

AU - Bisseling, T.

AU - Geurts, R.

PY - 2004

Y1 - 2004

N2 - Fluorescent proteins, such as green fluorescent protein and red fluorescent protein (DsRED), have become frequently used reporters in plant biology. However, their potential to monitor dynamic gene regulation is limited by their high stability. The recently made DsRED-E5 variant overcame this problem. DsRED-E5 changes its emission spectrum over time from green to red in a concentration independent manner. Therefore, the green to red fluorescence ratio indicates the age of the protein and can be used as a fluorescent timer to monitor dynamics of gene expression. Here, we analyzed the potential of DsRED-E5 as reporter in plant cells. We showed that in cowpea (Vigna unguiculata) mesophyll protoplasts, DsRED-E5 changes its fluorescence in a way similar to animal cells. Moreover, the timing of this shift is suitable to study developmental processes in plants. To test whether DsRed-E5 can be used to monitor gene regulation in plant organs, we placed DsRED-E5 under the control of promoters that are either up- or down-regulated (MtACT4 and LeEXT1. promoters) or constitutively expressed (MtACT2 promoter) during root hair development in Medicago truncatula. Analysis of the fluorescence ratios clearly provided more accurate insight into the timing of promoter activity.

AB - Fluorescent proteins, such as green fluorescent protein and red fluorescent protein (DsRED), have become frequently used reporters in plant biology. However, their potential to monitor dynamic gene regulation is limited by their high stability. The recently made DsRED-E5 variant overcame this problem. DsRED-E5 changes its emission spectrum over time from green to red in a concentration independent manner. Therefore, the green to red fluorescence ratio indicates the age of the protein and can be used as a fluorescent timer to monitor dynamics of gene expression. Here, we analyzed the potential of DsRED-E5 as reporter in plant cells. We showed that in cowpea (Vigna unguiculata) mesophyll protoplasts, DsRED-E5 changes its fluorescence in a way similar to animal cells. Moreover, the timing of this shift is suitable to study developmental processes in plants. To test whether DsRed-E5 can be used to monitor gene regulation in plant organs, we placed DsRED-E5 under the control of promoters that are either up- or down-regulated (MtACT4 and LeEXT1. promoters) or constitutively expressed (MtACT2 promoter) during root hair development in Medicago truncatula. Analysis of the fluorescence ratios clearly provided more accurate insight into the timing of promoter activity.

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