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A new design of a lentiviral shRNA vector with inducible co-expression of ARGONAUTE 2 for enhancing gene silencing efficiency

Overview of attention for article published in Cell & Bioscience, December 2015
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Title
A new design of a lentiviral shRNA vector with inducible co-expression of ARGONAUTE 2 for enhancing gene silencing efficiency
Published in
Cell & Bioscience, December 2015
DOI 10.1186/s13578-015-0058-2
Pubmed ID
Authors

Jiening He, Lian Huang, Huiling Qiu, Jiexuan Li, Lan Luo, Yanjiao Li, Shengli Tian, Kang Kang, Jun Luo, Lin Liu, Deming Gou

Abstract

RNA interference (RNAi) is a robust tool for inhibiting specific gene expression, but it is limited by the uncertain efficiency of siRNA or shRNA constructs. It has been shown that the overexpression of ARGONAUTE 2 (AGO2) protein increases silencing efficiency. However, the key elements required for AGO2-mediated enhancement of gene silencing in lentiviral vector has not been well studied. To explore the application of AGO2-based shRNA system in mammalian cells, we designed shRNA vectors targeting the EGFP reporter gene and evaluated the effects of various factors on silencing efficiency including stem length, loop sequence, antisense location as well as the ratio between AGO2 and shRNA. We found that 19 ~ 21-bp stem and 6- or 9-nt loop structure in the sense-loop-antisense (S-L-AS) orientation was an optimal design in the AGO2-shRNA system. Then, we constructed a single lentiviral vector co-expressing shRNA and AGO2 and demonstrated that the simultaneous expression of shRNA and AGO2 can achieve robust silencing of exogenous DsRed2 and endogenous ID1 and P65 genes. However, the titers of packaged lentivirus from constitutive expression of AGO2 vector were extremely low, severely limiting its broad application. For the first time, we demonstrated that the problem can be significantly improved by using the inducible expression of AGO2 lentiviral system. We reported a novel lentiviral vector with an optimal design of shRNA and inducible AGO2 overexpression which provides a new tool for RNAi research.

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Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 16 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
United States 1 6%
Unknown 15 94%

Demographic breakdown

Readers by professional status Count As %
Researcher 7 44%
Student > Ph. D. Student 2 13%
Student > Bachelor 2 13%
Lecturer > Senior Lecturer 1 6%
Unknown 4 25%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 5 31%
Agricultural and Biological Sciences 4 25%
Pharmacology, Toxicology and Pharmaceutical Science 1 6%
Neuroscience 1 6%
Chemistry 1 6%
Other 0 0%
Unknown 4 25%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 08 December 2015.
All research outputs
#18,432,465
of 22,835,198 outputs
Outputs from Cell & Bioscience
#559
of 930 outputs
Outputs of similar age
#280,659
of 388,741 outputs
Outputs of similar age from Cell & Bioscience
#16
of 19 outputs
Altmetric has tracked 22,835,198 research outputs across all sources so far. This one is in the 11th percentile – i.e., 11% of other outputs scored the same or lower than it.
So far Altmetric has tracked 930 research outputs from this source. They receive a mean Attention Score of 3.5. This one is in the 12th percentile – i.e., 12% of its peers scored the same or lower than it.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 388,741 tracked outputs that were published within six weeks on either side of this one in any source. This one is in the 16th percentile – i.e., 16% of its contemporaries scored the same or lower than it.
We're also able to compare this research output to 19 others from the same source and published within six weeks on either side of this one. This one is in the 1st percentile – i.e., 1% of its contemporaries scored the same or lower than it.