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Related Concept Videos

Reporter Genes02:11

Reporter Genes

Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
Commonly used reporter...
Labeling DNA Probes03:31

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
Ribosome Profiling02:24

Ribosome Profiling

Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...

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Massively Parallel Reporter Assays in Cultured Mammalian Cells
11:03

Massively Parallel Reporter Assays in Cultured Mammalian Cells

Published on: August 17, 2014

RUBY-BD: a highly efficient visual reporter system for assessing bidirectional promoter activity.

Qi Sha1, Wen Zhou1, XinYan Zhao1

  • 1College of Agriculture and Biology, Liaocheng University, Liaocheng, 252000, China.

Planta
|May 27, 2026
PubMed
Summary
This summary is machine-generated.

Researchers created RUBY-BD, a novel reporter system for high-throughput analysis of bidirectional promoters (BDPs). This visual tool simplifies BDP characterization, accelerating plant gene engineering and research.

Keywords:
RUBY-BDBetalainsBidirectional promoterMultigene engineeringTranscription activity

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Area of Science:

  • Molecular Biology
  • Plant Science
  • Biotechnology

Background:

  • Bidirectional promoters (BDPs) are crucial for simultaneous transcription of adjacent genes, valuable in multigene engineering.
  • Existing reporter systems (GFP, GUS) for BDP activity assessment are costly and labor-intensive, limiting high-throughput screening.
  • The RUBY reporter system utilizes enzymes to convert tyrosine into betalain, a visually detectable pigment.

Purpose of the Study:

  • To develop a universal, one-step reporter system for high-throughput examination of plant bidirectional promoter activity.
  • To construct and validate the RUBY-BD vector for efficient screening of BDPs.
  • To accelerate the discovery and application of BDPs in plant gene engineering.

Main Methods:

  • Development of the RUBY-BD vector by separating RUBY reporter genes into two bidirectional transcription units.
  • Transient transformation assays in Nicotiana benthamiana leaves to test BDP activity.
  • Visual assessment of betalain accumulation and expression analysis of flanking genes to confirm BDP function.

Main Results:

  • The RUBY-BD system successfully enabled betalain biosynthesis upon insertion of active BDPs.
  • Evaluation of 20 putative plant BDPs using the RUBY-BD system in a single step.
  • Confirmation of bidirectional transcriptional activity in 11 out of 20 tested BDPs.

Conclusions:

  • The RUBY-BD system provides a high-throughput, visual screening tool for characterizing plant BDPs.
  • This system simplifies the assessment of BDP activity, overcoming limitations of conventional methods.
  • The RUBY-BD system is expected to significantly advance basic research and applications of BDPs in plant biotechnology.