Dna Reporter Construct
Dna Reporter Construct - Related Videos
07:19
Identifying Targets of Human microRNAs with the LightSwitch Luciferase Assay System using 3'UTR-reporter Constructs and a microRNA Mimic in Adherent Cells
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Cited by 22 •
2011
MicroRNAs (miRNAs) are important regulators of gene expression and have been shown to play a role in numerous biological processes. To better understand miRNA-UTR interactions, we have created a genome-wide collection of 3 UTR luciferase reporters paired with a novel luciferase gene and assay reagent, the LightSwitch system.
09:32
Stable DNA Motifs, 1D and 2D Nanostructures Constructed from Small Circular DNA Molecules
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Cited by 6 •
2019
This article presents a detailed protocol for T4 ligation and denaturing PAGE purification of small circular DNA molecules, annealing and native PAGE analysis of circular tiles, assembling and AFM imaging of 1D and 2D DNA nanostructures, as well as agarose gel electrophoresis and centrifugation purification of finite DNA nanostructures.
12:59
High-throughput CRISPR Vector Construction and Characterization of DNA Modifications by Generation of Tomato Hairy Roots
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Cited by 37 •
2016
Using DNA assembly, multiple CRISPR vectors can be constructed in parallel in a single cloning reaction, making the construction of large numbers of CRISPR vectors a simple task. Tomato hairy roots are an excellent model system to validate CRISPR vectors and generate mutant materials.
12:48
Construction of Cell-based Neurotransmitter Fluorescent Engineered Reporters (CNiFERs) for Optical Detection of Neurotransmitters In Vivo
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Cited by 17 •
2016
We present a protocol to create cell-based neurotransmitter fluorescent engineered reporters (CNiFERs) for the optical detection of volumetric neurotransmitter release.
11:03
Massively Parallel Reporter Assays in Cultured Mammalian Cells
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Cited by 63 •
2014
The genetic reporter assay is a well-established and powerful tool for dissecting the relationship between DNA sequences and their gene regulatory activities. Coupling candidate regulatory elements to reporter genes that carry identifying sequence tags enables massive parallelization of these assays.
FAQs
The regulatory sequence determines which biological activity is represented by the reporter signal. Linking a promoter or other regulatory element to the reporter allows researchers to examine whether that element becomes active under a particular cellular condition. Consequently, the sequence selected for the construct should match the regulatory event or gene expression process being investigated.
The reporter gene converts regulatory activity into an observable output. Cellular machinery transcribes and translates the reporter, producing fluorescence, luminescence, or an enzymatic color change. Because these outputs can be detected visually or measured quantitatively, the reporter connects an otherwise molecular event to an experimental readout of gene expression or pathway activity.
A detectable reporter signal depends on the information in the construct being expressed through cellular machinery. Activation of the regulatory element initiates transcription of the reporter, and translation produces the reporter product responsible for the observable output. This two-stage mechanism allows the construct to link regulatory control with measurable evidence of cellular activity.
These outputs provide different ways to observe the same underlying regulatory event. Fluorescence can provide a visual signal, luminescence can serve as a measurable output, and an enzymatic reporter can produce a color change. The choice among them affects whether the experiment emphasizes visual evidence, quantitative measurement, or an observable enzymatic response.
Researchers first link the regulatory sequence or gene of interest to a reporter gene, then use the engineered DNA in an experimental system such as living cells. After the relevant regulatory element is activated, cellular transcription and translation generate the reporter output. The resulting fluorescence, luminescence, or color change is then used to assess the biological event.
Reporter constructs can be applied to questions about promoter function, transcriptional regulation, cellular responses, and pathway activity. They provide a way to connect these processes with visible or measurable outputs rather than relying only on the underlying DNA sequence. In living cells and other experimental systems, this supports analysis of molecular events through quantitative or visual evidence.
A reporter output can provide detectable evidence in experiments involving gene delivery. When the delivered construct reaches an experimental system and its regulatory element becomes active, reporter expression can produce fluorescence, luminescence, or an enzymatic color change. This makes the construct useful for examining whether delivered genetic information leads to an observable molecular response.
The signal can indicate whether a regulatory element is active and can provide either quantitative or visual evidence of gene expression, pathway activity, or a cellular response. Interpretation depends on the reporter output and the biological question. In suitable experimental systems, the measured or observed signal connects molecular regulation with a detectable experimental outcome.