Studying Transcription Factor Activity with the LightSwitch System

A SOLUTION FOR:

Monitoring changes in promoter activity in response to variations in transcription factor function
Assessing the functional consequences of transcription factor binding detected through a ChIP-based experiment
Measuring the effects of mutagenizing a putative transcription factor binding motif
Verifying computational predictions and supplementing microarray or next generation sequencing data

ADVANTAGES:

Optimal performance: Novel RenSP luciferase technology allows you to measure promoter or 3’UTR activity with industry-leading sensitivity and dynamic range
Simple, quick, complete solution: Complete LightSwitch Luciferase Assay System enables easy transfection of luciferase reporter constructs and controls followed by detecting reporter signal with optimized LightSwitch Luciferase Assay Reagents
Comprehensive and verified: Choose from a human genome-wide collection of 30,000 transfection-ready sequence-verified promoter and 3'UTR reporter constructs. No cloning, DNA preparation, or reagent optimization is needed

OVERVIEW

Transcription factors are important regulators of eukaryotic transcriptional regulation, and they interact with DNA-encoded regulatory elements like promoters to modulate when, where, and how much of a gene’s mRNA gets made. The activity of transcription factors can be affected by the cell’s environmental conditions including exposure to signaling molecules like hormones and growth factors. Reporter assays are a powerful technique for measuring the activity of promoters in living cells. GoClone Reporter Constructs are created by isolating a promoter from the human genome and cloning it upstream of the RenSP luciferase reporter gene on a plasmid. Once this plasmid is transfected into a living cell, a change in promoter activity causes a change in reporter signal (light output).

The LightSwitch Luciferase Assay System and Promoter Reporter GoClone Collection are important tools for determining how a transcription factor regulates its target genes. Researchers can monitor changes in any promoter’s activity in response to variations in transcription factor function that may be modulated by techniques like over-expression (often by cDNA co-transfection), knock-down (often by siRNA co-transfection), or treatment with a known agonist or antagonist. The Promoter Reporter GoClones are also useful for assessing the functional consequences of transcription factor binding detected through a ChIP-based experiment and for measuring the effects of mutagenizing a putative transcription factor binding motif.

RESOURCES
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