Expression of Multiple Artificial MicroRNAs from a Chicken miRNA126-Based Lentiviral Vector
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Chen-2011-Expression of Multip.pdf
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Author(s)
Chen, Steve C.-Y.
Stern, Patrick
Guo, Zhuyan
Chen, Jianzhu
Date Issued
July 2011
Journal
PLoS ONE
Publisher
Public Library of Science
Citation
Chen, Steve C.-Y. et al. “Expression of Multiple Artificial MicroRNAs from a Chicken miRNA126-Based Lentiviral Vector.” Ed. Lennart Randau. PLoS ONE 6 (7) (2011): e22437. © 2011 Chen et al.
Version
Final published version
Abstract
Background: The use of RNAi in both basic and translational research often requires expression of multiple siRNAs from the
same vector.
Methods/Principal Findings: We have developed a novel chicken miR126-based artificial miRNA expression system that can
express one, two or three miRNAs from a single cassette in a lentiviral vector. We show that each of the miRNAs expressed
from the same lentiviral vector is capable of potent inhibition of reporter gene expression in transient transfection and
stable integration assays in chicken fibroblast DF-1 cells. Transduction of Vero cells with lentivirus expressing two or three
different anti-influenza miRNAs leads to inhibition of influenza virus production. In addition, the chicken miR126-based
expression system effectively inhibits reporter gene expression in human, monkey, dog and mouse cells. These results
demonstrate that the flanking regions of a single primary miRNA can support processing of three different stem-loops in a
single vector.
Conclusions/Significance: This novel design expands the means to express multiple miRNAs from the same vector for
potent and effective silencing of target genes and influenza virus.
MIT Department
Massachusetts Institute of Technology. Department of Biology
Koch Institute for Integrative Cancer Research at MIT
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DOI of Published Version
https://doi.org/10.1371/journal.pone.0022437
