High-Throughput Functional MicroRNAs Profiling by Recombinant AAV-Based MicroRNA Sensor Arrays

Tian, Wenhong; Dong, Xiaoyan; Liu, Xuerong; Wang, Gang; Dong, Zheyue; Shen, Wei; Zheng, Gang; Lü, Jianxin; Chen, Jinzhong; Wang, Yue; Wu, Zhijian; Wu, Xiaobing

doi:10.1371/journal.pone.0029551

Cited by 47 publications

(54 citation statements)

References 45 publications

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“…In Huh7 cells,which are hepatocellular carcinoma well-known to express high levels of endogenous miR-122, [16] we obtained unequivocal results showing successful and simultaneous release/inhibition of miR-122 function by both Ant122 and sm122 (Steps V, VI), and leading to combination inhibition of endogenous miR-122 function.…”

Section: Angewandte Communicationsmentioning

confidence: 81%

Single‐Vehicular Delivery of Antagomir and Small Molecules to Inhibit miR‐122 Function in Hepatocellular Carcinoma Cells by using “Smart” Mesoporous Silica Nanoparticles

Qian

Uttamchandani

et al. 2015

Angew Chem Int Ed

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MicroRNAs (miRNAs) regulate a variety of biological processes. The liver-specific, highly abundant miR-122 is implicated in many human diseases including cancer. Its inhibition has been found to result in a dramatic loss in the ability of Hepatitis C virus (HCV) to infect host cells. Both antisense technology and small molecules have been used to independently inhibit endogenous miR-122 function, but not in combination. Intracellular stability, efficient delivery, hydrophobicity, and controlled release are some of the current challenges associated with these novel therapeutic methods. Reported herein is the first single-vehicular system, based on mesoporous silica nanoparticles (MSNs), for simultaneous cellular delivery of miR-122 antagomir and small molecule inhibitors. The controlled release of both types of inhibitors depends on the expression levels of endogenous miR-122, thus enabling these drug-loaded MSNs to achieve combination inhibition of its targeted mRNAs in Huh7 cells.

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Section: Angewandte Communicationsmentioning

confidence: 81%

Single‐Vehicular Delivery of Antagomir and Small Molecules to Inhibit miR‐122 Function in Hepatocellular Carcinoma Cells by using “Smart” Mesoporous Silica Nanoparticles

Qian

Uttamchandani

et al. 2015

Angew Chem Int Ed

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“…At least two sensor library systems to quantify miRNA activity in a high-throughput manner have been developed in mammals (Mullokandov et al, 2012;Tian et al, 2012). These systems can be used for monitoring miRNA activities in response to various stimuli, such as chemical compounds or environmental conditions.…”

Section: Discussionmentioning

confidence: 99%

Dissection of miRNA Pathways Using Arabidopsis Mesophyll Protoplasts

et al. 2015

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MicroRNAs (miRNAs) control gene expression mostly post-transcriptionally by guiding transcript cleavage and/or translational repression of complementary mRNA targets, thereby regulating developmental processes and stress responses. Despite the remarkable expansion of the field, the mechanisms underlying miRNA activity are not fully understood. In this article, we describe a transient expression system in Arabidopsis mesophyll protoplasts, which is highly amenable for the dissection of miRNA pathways. We show that by transiently overexpressing primary miRNAs and target mimics, we can manipulate miRNA levels and consequently impact on their targets. Furthermore, we developed a set of luciferase-based sensors for quantifying miRNA activity that respond specifically to both endogenous and overexpressed miRNAs and target mimics. We demonstrate that these miRNA sensors can be used to test the impact of putative components of the miRNA pathway on miRNA activity, as well as the impact of specific mutations, by either overexpression or the use of protoplasts from the corresponding mutants. We further show that our miRNA sensors can be used for investigating the effect of chemicals on miRNA activity. Our cell-based transient expression system is fast and easy to set up, and generates quantitative results, being a powerful tool for assaying miRNA activity in vivo.

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“…Recently, at least two sensor library systems to quantify miRNA activity in a high throughput manner have been developed in mammals (Mullokandov et al, 2012;Tian et al, 2012). These systems can be used for monitoring miRNA activities in response to various stimuli, such as chemical compounds or environmental conditions.…”

Section: Discussionmentioning

confidence: 99%

Dissection of miRNA pathways using Arabidopsis mesophyll protoplasts

Martinho¹,

Confraria²,

Elias³

et al. 2014

Molecular Plant

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Short summary: We developed a set of sensors based on the firefly luciferase reporter gene to quantify miRNA activity in vivo, using Arabidopsis mesophyll protoplasts. We show that these sensors are not only responsive to endogenous miRNA activity, but also to exogenous gain-and loss-of-function manipulation of miRNA abundance. As examples, our sensors can be used for studying regulatory mechanisms of miRNA-associated components, for identifying novel factors with miRNA-associated functions, and for assessing the effect of chemicals on miRNA activity, therefore constituting a powerful tool to dissect miRNA pathways. responses. Despite the remarkable expansion of the field, the mechanisms underlying miRNA activity are not fully understood. In this paper, we describe a transient expression system in Arabidopsis mesophyll protoplasts that is highly amenable for the dissection of miRNA pathways. We show that by transiently overexpressing primary miRNAs and target mimics, we can manipulate miRNA levels and consequently impact on their targets. Furthermore, we developed a set of luciferase-based sensors for quantifying miRNA activity that respond specifically to both endogenous and overexpressed miRNAs and target mimics.We demonstrate that these miRNA sensors can be used to test the impact of putative components of the miRNA pathway on miRNA activity, as well as the impact of specific mutations, either by overexpression or by the use of protoplasts from the corresponding mutants. We further show that our miRNA sensors can be used for investigating the effect of chemicals on miRNA activity.Our cell-based transient expression system is fast and easy to set up and generates quantitative results, being a powerful tool for assaying miRNA activity in vivo.

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High-Throughput Functional MicroRNAs Profiling by Recombinant AAV-Based MicroRNA Sensor Arrays

Cited by 47 publications

References 45 publications

Single‐Vehicular Delivery of Antagomir and Small Molecules to Inhibit miR‐122 Function in Hepatocellular Carcinoma Cells by using “Smart” Mesoporous Silica Nanoparticles

Single‐Vehicular Delivery of Antagomir and Small Molecules to Inhibit miR‐122 Function in Hepatocellular Carcinoma Cells by using “Smart” Mesoporous Silica Nanoparticles

Dissection of miRNA Pathways Using Arabidopsis Mesophyll Protoplasts

Dissection of miRNA pathways using Arabidopsis mesophyll protoplasts

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