Designing Ago2-specific siRNA/shRNA to Avoid Competition with Endogenous miRNAs
Relatively large amounts of transfected siRNA can compete for Ago proteins and thus compromise endogenous miRNA function, potentially leading to toxicities. Here, we show that shRNA can also perturb endogenous miRNA function similarly. More importantly, we also show that the problem can be solved by...
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2014-01-01
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Online Access: | http://www.sciencedirect.com/science/article/pii/S216225311630316X |
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doaj-b25285ffd1734cfd9edfb64268b7beec2020-11-24T23:12:23ZengElsevierMolecular Therapy: Nucleic Acids2162-25312014-01-013C10.1038/mtna.2014.27Designing Ago2-specific siRNA/shRNA to Avoid Competition with Endogenous miRNAsHongming Ma0Junli Zhang1Haoquan Wu2Department of Biomedical Sciences, Center of Excellence in Infectious Disease Research, Paul L. Foster School of Medicine, Texas Tech University Health Sciences Center, El Paso, Texas, USADepartment of Biomedical Sciences, Center of Excellence in Infectious Disease Research, Paul L. Foster School of Medicine, Texas Tech University Health Sciences Center, El Paso, Texas, USADepartment of Biomedical Sciences, Center of Excellence in Infectious Disease Research, Paul L. Foster School of Medicine, Texas Tech University Health Sciences Center, El Paso, Texas, USARelatively large amounts of transfected siRNA can compete for Ago proteins and thus compromise endogenous miRNA function, potentially leading to toxicities. Here, we show that shRNA can also perturb endogenous miRNA function similarly. More importantly, we also show that the problem can be solved by designing shRNAs in the context of pre-miR-451 structure with completely complementary stem, which significantly improves the Ago2 specificity. This shRNA was shown to be Ago2-specific, and maintain target-silencing ability while avoiding competition with endogenous miRNAs by not competing for Agos 1, 3, and 4. We conclude that modified pre-miR-451 structure provides a general platform to design shRNAs that significantly reduce perturbation of miRNA function.http://www.sciencedirect.com/science/article/pii/S216225311630316XAgo2DicermiR-451shRNA |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Hongming Ma Junli Zhang Haoquan Wu |
spellingShingle |
Hongming Ma Junli Zhang Haoquan Wu Designing Ago2-specific siRNA/shRNA to Avoid Competition with Endogenous miRNAs Molecular Therapy: Nucleic Acids Ago2 Dicer miR-451 shRNA |
author_facet |
Hongming Ma Junli Zhang Haoquan Wu |
author_sort |
Hongming Ma |
title |
Designing Ago2-specific siRNA/shRNA to Avoid Competition with Endogenous miRNAs |
title_short |
Designing Ago2-specific siRNA/shRNA to Avoid Competition with Endogenous miRNAs |
title_full |
Designing Ago2-specific siRNA/shRNA to Avoid Competition with Endogenous miRNAs |
title_fullStr |
Designing Ago2-specific siRNA/shRNA to Avoid Competition with Endogenous miRNAs |
title_full_unstemmed |
Designing Ago2-specific siRNA/shRNA to Avoid Competition with Endogenous miRNAs |
title_sort |
designing ago2-specific sirna/shrna to avoid competition with endogenous mirnas |
publisher |
Elsevier |
series |
Molecular Therapy: Nucleic Acids |
issn |
2162-2531 |
publishDate |
2014-01-01 |
description |
Relatively large amounts of transfected siRNA can compete for Ago proteins and thus compromise endogenous miRNA function, potentially leading to toxicities. Here, we show that shRNA can also perturb endogenous miRNA function similarly. More importantly, we also show that the problem can be solved by designing shRNAs in the context of pre-miR-451 structure with completely complementary stem, which significantly improves the Ago2 specificity. This shRNA was shown to be Ago2-specific, and maintain target-silencing ability while avoiding competition with endogenous miRNAs by not competing for Agos 1, 3, and 4. We conclude that modified pre-miR-451 structure provides a general platform to design shRNAs that significantly reduce perturbation of miRNA function. |
topic |
Ago2 Dicer miR-451 shRNA |
url |
http://www.sciencedirect.com/science/article/pii/S216225311630316X |
work_keys_str_mv |
AT hongmingma designingago2specificsirnashrnatoavoidcompetitionwithendogenousmirnas AT junlizhang designingago2specificsirnashrnatoavoidcompetitionwithendogenousmirnas AT haoquanwu designingago2specificsirnashrnatoavoidcompetitionwithendogenousmirnas |
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1725600965000691712 |