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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...
MicroRNAs01:22

MicroRNAs

MicroRNA (miRNA) are short, regulatory RNA transcribed from introns—non-coding regions of a gene—or intergenic regions—stretches of DNA present between genes. Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA ends...
MicroRNAs01:22

MicroRNAs

MicroRNA (miRNA) are short, regulatory RNA transcribed from introns (non-coding regions of a gene) or intergenic regions (stretches of DNA present between genes). Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself, forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA...

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Related Experiment Video

Updated: Jun 29, 2026

In ovo Expression of MicroRNA in Ventral Chick Midbrain
09:19

In ovo Expression of MicroRNA in Ventral Chick Midbrain

Published on: September 16, 2013

A construct with fluorescent indicators for conditional expression of miRNA.

Linghua Qiu1, Hongyan Wang, Xugang Xia

  • 1Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, 364 Plantation St, Worcester, MA 01605, USA. Linghua.Qiu@umassmed.edu

BMC Biotechnology
|October 9, 2008
PubMed
Summary

We developed an improved method for creating transgenic animals using inducible RNA interference (RNAi) and fluorescent markers. This approach enhances gene silencing efficiency and simplifies the analysis of genetically modified organisms.

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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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Identifying Targets of Human microRNAs with the LightSwitch Luciferase Assay System using 3'UTR-reporter Constructs and a microRNA Mimic in Adherent Cells

Published on: September 28, 2011

A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells
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A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells

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Related Experiment Videos

Last Updated: Jun 29, 2026

In ovo Expression of MicroRNA in Ventral Chick Midbrain
09:19

In ovo Expression of MicroRNA in Ventral Chick Midbrain

Published on: September 16, 2013

Identifying Targets of Human microRNAs with the LightSwitch Luciferase Assay System using 3'UTR-reporter Constructs and a microRNA Mimic in Adherent Cells
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

Published on: September 28, 2011

A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells
06:48

A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells

Published on: June 16, 2022

Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Transgenic RNA interference (RNAi) offers a cost-effective method for reverse genetics in mammals.
  • Inducible RNAi allows for controlled gene silencing in vivo, enhancing its utility.
  • Simultaneous expression of microRNA (miRNA) and fluorescent markers aids in screening transgenic animals.

Purpose of the Study:

  • To optimize the simultaneous expression of miRNA and fluorescent markers in transgenic systems.
  • To compare construct designs for efficient miRNA and reporter protein expression.
  • To develop a conditional expression system for inducible gene silencing.

Main Methods:

  • Compared two construct designs: one with miRNA in the 3' untranslated region (UTR) and another with the precursor miRNA (pre-miRNA) in an intron within the 3'-UTR.
  • Assessed miRNA and red fluorescent protein (RFP) expression levels.
  • Developed a conditional construct using the Cre-loxP system for inducible expression.

Main Results:

  • The intron-containing construct significantly enhanced RFP expression compared to the intron-less design.
  • The intron improved gene expression by its own activity and by mitigating pre-miRNA inhibition.
  • The conditional construct successfully silenced EGFP and induced RFP and miRNA expression upon Cre recombinase exposure.

Conclusions:

  • A novel conditional miRNA-expression construct was designed and validated.
  • The construct strongly expresses marker genes and efficiently silences target genes upon induction.
  • This system improves the efficiency of generating cell lines and transgenic animals for miRNA-based gene silencing.