A Novel Transgenic Mouse Line for Tracing MicroRNA-155-5p Activity In Vivo

MicroRNA-155 (miR-155) plays significant role in various physiological processes involving both innate and adaptive immunity. miR-155 expression level changes dynamically during various immune responses. However, current approaches for miR-155 detection at the RNA level do not precisely reflect the...

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Bibliographic Details
Published in:PloS one 2015-06, Vol.10 (6), p.e0128198-e0128198
Main Authors: Phiwpan, Krung, Guo, Jie, Zhang, Wei, Hu, Tanyu, Boruah, Bhargavi M, Zhang, Jianhua, Zhou, Xuyu
Format: Article
Language:English
Subjects:
3' Untranslated Regions - genetics
Adaptive immunity
Animals
Antigens
Artificial chromosomes
B cells
Brain research
Cell Lineage
Chromosomes, Artificial, Bacterial - genetics
Cloning
Diphtheria
Diphtheria toxin
Diphtheria Toxin - genetics
Fluorescence
Gene expression
Genes, Reporter - genetics
Genetic engineering
Genetic Engineering - methods
Immune response
Immunity
Immunology
In vivo methods and tests
Laboratories
Luminescent Proteins - genetics
Lymphocytes
Mice
Mice, Transgenic
MicroRNA
MicroRNAs
MicroRNAs - genetics
miRNA
Physiological aspects
Polymerase chain reaction
Proteins
Ribonucleic acid
RNA
RNA polymerase
Rodents
Sensors
Transgenic animals
Transgenic mice
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Summary:MicroRNA-155 (miR-155) plays significant role in various physiological processes involving both innate and adaptive immunity. miR-155 expression level changes dynamically during various immune responses. However, current approaches for miR-155 detection at the RNA level do not precisely reflect the real-time activity. Herein, we generated a transgenic mouse line (R26-DTR-155T) for determination of miR-155-5p activity in vivo by inserting miR-155-5p target sequence downstream of a reporter transgene comprising Diphtheria Toxin Receptor and TagBlue fluorescence protein. Using this approach, R26-DTR-155T mice were able to measure variation in levels of miR-155-5p activity in specific cell types of interest. The DTR expression levels were inversely correlated with the endogenous miR-155 expression pattern as detected by quantitative RT-PCR. Our data demonstrate a novel transgenic mouse line which could be useful for tracing miR-155-5p activity in specific cell types through measurement of miR-155-5p activity at single cell level.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0128198