Small RNA Expression Profiling by High-Throughput Sequencing: Implications of Enzymatic Manipulation

Eukaryotic regulatory small RNAs (sRNAs) play significant roles in many fundamental cellular processes. As such, they have emerged as useful biomarkers for diseases and cell differentiation states. sRNA-based biomarkers outperform traditional messenger RNA-based biomarkers by testing fewer targets...

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Main Authors: Fanglei Zhuang, Ryan T. Fuchs, G. Brett Robb
Format: Article
Language:English
Published: Wiley 2012-01-01
Series:Journal of Nucleic Acids
Online Access:http://dx.doi.org/10.1155/2012/360358
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author Fanglei Zhuang
Ryan T. Fuchs
G. Brett Robb
author_facet Fanglei Zhuang
Ryan T. Fuchs
G. Brett Robb
author_sort Fanglei Zhuang
collection DOAJ
description Eukaryotic regulatory small RNAs (sRNAs) play significant roles in many fundamental cellular processes. As such, they have emerged as useful biomarkers for diseases and cell differentiation states. sRNA-based biomarkers outperform traditional messenger RNA-based biomarkers by testing fewer targets with greater accuracy and providing earlier detection for disease states. Therefore, expression profiling of sRNAs is fundamentally important to further advance the understanding of biological processes, as well as diagnosis and treatment of diseases. High-throughput sequencing (HTS) is a powerful approach for both sRNA discovery and expression profiling. Here, we discuss the general considerations for sRNA-based HTS profiling methods from RNA preparation to sequencing library construction, with a focus on the causes of systematic error. By examining the enzymatic manipulation steps of sRNA expression profiling, this paper aims to demystify current HTS-based sRNA profiling approaches and to aid researchers in the informed design and interpretation of profiling experiments.
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spelling doaj-art-abf94485bc81469381a4004008572d802025-02-03T05:44:02ZengWileyJournal of Nucleic Acids2090-02012090-021X2012-01-01201210.1155/2012/360358360358Small RNA Expression Profiling by High-Throughput Sequencing: Implications of Enzymatic ManipulationFanglei Zhuang0Ryan T. Fuchs1G. Brett Robb2RNA Division, New England Biolabs Inc., 240 County Road, Ipswich, MA 01938, USARNA Division, New England Biolabs Inc., 240 County Road, Ipswich, MA 01938, USARNA Division, New England Biolabs Inc., 240 County Road, Ipswich, MA 01938, USAEukaryotic regulatory small RNAs (sRNAs) play significant roles in many fundamental cellular processes. As such, they have emerged as useful biomarkers for diseases and cell differentiation states. sRNA-based biomarkers outperform traditional messenger RNA-based biomarkers by testing fewer targets with greater accuracy and providing earlier detection for disease states. Therefore, expression profiling of sRNAs is fundamentally important to further advance the understanding of biological processes, as well as diagnosis and treatment of diseases. High-throughput sequencing (HTS) is a powerful approach for both sRNA discovery and expression profiling. Here, we discuss the general considerations for sRNA-based HTS profiling methods from RNA preparation to sequencing library construction, with a focus on the causes of systematic error. By examining the enzymatic manipulation steps of sRNA expression profiling, this paper aims to demystify current HTS-based sRNA profiling approaches and to aid researchers in the informed design and interpretation of profiling experiments.http://dx.doi.org/10.1155/2012/360358
spellingShingle Fanglei Zhuang
Ryan T. Fuchs
G. Brett Robb
Small RNA Expression Profiling by High-Throughput Sequencing: Implications of Enzymatic Manipulation
Journal of Nucleic Acids
title Small RNA Expression Profiling by High-Throughput Sequencing: Implications of Enzymatic Manipulation
title_full Small RNA Expression Profiling by High-Throughput Sequencing: Implications of Enzymatic Manipulation
title_fullStr Small RNA Expression Profiling by High-Throughput Sequencing: Implications of Enzymatic Manipulation
title_full_unstemmed Small RNA Expression Profiling by High-Throughput Sequencing: Implications of Enzymatic Manipulation
title_short Small RNA Expression Profiling by High-Throughput Sequencing: Implications of Enzymatic Manipulation
title_sort small rna expression profiling by high throughput sequencing implications of enzymatic manipulation
url http://dx.doi.org/10.1155/2012/360358
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