Small RNA Sequencing across Diverse Biofluids Identifies Optimal Methods for exRNA Isolation

Poor reproducibility within and across studies arising from lack of knowledge regarding the performance of extracellular RNA (exRNA) isolation methods has hindered progress in the exRNA field. A systematic comparison of 10 exRNA isolation methods across 5 biofluids revealed marked differences in the...

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Published in:Cell Vol. 177; no. 2; p. 446
Main Authors: Srinivasan, Srimeenakshi, Yeri, Ashish, Cheah, Pike See, Chung, Allen, Danielson, Kirsty, De Hoff, Peter, Filant, Justyna, Laurent, Clara D, Laurent, Lucie D, Magee, Rogan, Moeller, Courtney, Murthy, Venkatesh L, Nejad, Parham, Paul, Anu, Rigoutsos, Isidore, Rodosthenous, Rodosthenis, Shah, Ravi V, Simonson, Bridget, To, Cuong, Wong, David, Yan, Irene K, Zhang, Xuan, Balaj, Leonora, Breakefield, Xandra O, Daaboul, George, Gandhi, Roopali, Lapidus, Jodi, Londin, Eric, Patel, Tushar, Raffai, Robert L, Sood, Anil K, Alexander, Roger P, Das, Saumya, Laurent, Louise C
Format: Journal Article
Language:English
Published: United States 04.04.2019
Subjects:
Adult
Body Fluids - chemistry
Cell Line
Cell-Free Nucleic Acids - isolation & purification
Circulating MicroRNA - isolation & purification
Extracellular Vesicles - metabolism
Female
Healthy Volunteers
Humans
Male
MicroRNAs - isolation & purification
MicroRNAs - metabolism
Reproducibility of Results
RNA - isolation & purification
RNA - metabolism
Sequence Analysis, RNA - methods
extracellular vesicles
lipoprotein
ribonucleoprotein
extracellular RNA
ISSN:1097-4172, 1097-4172
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Summary:Poor reproducibility within and across studies arising from lack of knowledge regarding the performance of extracellular RNA (exRNA) isolation methods has hindered progress in the exRNA field. A systematic comparison of 10 exRNA isolation methods across 5 biofluids revealed marked differences in the complexity and reproducibility of the resulting small RNA-seq profiles. The relative efficiency with which each method accessed different exRNA carrier subclasses was determined by estimating the proportions of extracellular vesicle (EV)-, ribonucleoprotein (RNP)-, and high-density lipoprotein (HDL)-specific miRNA signatures in each profile. An interactive web-based application (miRDaR) was developed to help investigators select the optimal exRNA isolation method for their studies. miRDar provides comparative statistics for all expressed miRNAs or a selected subset of miRNAs in the desired biofluid for each exRNA isolation method and returns a ranked list of exRNA isolation methods prioritized by complexity, expression level, and reproducibility. These results will improve reproducibility and stimulate further progress in exRNA biomarker development.
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ISSN:1097-4172
1097-4172
DOI:10.1016/j.cell.2019.03.024