scholarly journals Evaluation of Commercially Available Exosomal Isolation Kits from Human Plasma

2019 ◽  
Author(s):  
Yuzhe Sun ◽  
Hefu Zhen ◽  
Mei Guo ◽  
Jingyu Ye ◽  
Zhili Liu ◽  
...  

AbstractExosomes are cell-derived lipid bilayer particles which are abundant in biological fluids. Exosome is an emerging source of biomarkers to diagnose various human diseases. Sequencing based exosomal studies could provide a comprehensive view of exosomal RNA and protein. To extracted these inclusions, exosomes should be isolated from the plasma first. Several exosome isolation methods were introduced since the discover of exosome. To promote the clinical application of exosomal inclusions, different isolation methods should be compared. We isolated exosomes from human plasma by using user-friendly and commercially available kits, SBI ExoQuick and QIAGEN exoRNeasy. Subsequently, small RNA sequencing was performed with two groups of isolated exosome samples and one group of plasma samples. No fundamental differences of exRNA yield between SC and EQ were found. In RNA profile analysis, the small RNA aligned reads, miRNA pattern, sample clustering varied as a result of methodological differences. Small RNA isolated by ExoQuick presented better data quality and RNA profile than exoRNeasy. This study compared sRNA sequencing data generated from two exosome isolation kits, it provides a reference for future small RNA studies and biomarker prediction in human plasma exosome.

Genes ◽  
2021 ◽  
Vol 12 (10) ◽  
pp. 1576
Author(s):  
Jin-Ok Lee ◽  
Minho Lee ◽  
Yeun-Jun Chung

Transfer RNA (tRNA), a key component of the translation machinery, plays critical roles in stress conditions and various diseases. While knowledge regarding the importance of tRNA function is increasing, its biological roles are still not well understood. There is currently no comprehensive database or web server providing the expression landscape of tRNAs across a variety of human tissues and diseases. Here, we constructed a user-friendly and interactive database, DBtRend, which provides a profile of mature tRNA expression across various biological conditions by reanalyzing the small RNA or microRNA sequencing data from the Cancer Genome Atlas (TCGA) and NCBI’s Gene Expression Omnibus (GEO) in humans. Users can explore not only the expression values of mature individual tRNAs in the human genome, but also those of isodecoders and isoacceptors based on our specific pipelines. DBtRend provides the expressed patterns of tRNAs, the differentially expressed tRNAs in different biological conditions, and the information of samples or patients, tissue types, and molecular subtype of cancers. The database is expected to help researchers interested in functional discoveries of tRNAs.


2013 ◽  
Vol 35 (4) ◽  
pp. 342-347 ◽  
Author(s):  
Jeongsoo Lee ◽  
Dong-in Kim ◽  
June Hyun Park ◽  
Ik-Young Choi ◽  
Chanseok Shin

2016 ◽  
Author(s):  
Guillaume Carissimo ◽  
Marius van den Beek ◽  
Juliana Pegoraro ◽  
Kenneth D Vernick ◽  
Christophe Antoniewski

AbstractWe present user-friendly and adaptable software to provide biologists, clinical researchers and possibly diagnostic clinicians with the ability to robustly detect and reconstruct viral genomes from complex deep sequence datasets. A set of modular bioinformatic tools and workflows was implemented as the Metavisitor package in the Galaxy framework. Using the graphical Galaxy workflow editor, users with minimal computational skills can use existing Metavisitor workflows or adapt them to suit specific needs by adding or modifying analysis modules. Metavisitor can be used on our Mississippi server, or can be installed on any Galaxy server instance and a pre-configured Metavisitor server image is provided. Metavisitor works with DNA, RNA or small RNA sequencing data over a range of read lengths and can use a combination of de novo and guided approaches to assemble genomes from sequencing reads. We show that the software has the potential for quick diagnosis as well as discovery of viruses from a vast array of organisms. Importantly, we provide here executable Metavisitor use cases, which increase the accessibility and transparency of the software, ultimately enabling biologists or clinicians to focus on biological or medical questions.


Plants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 267
Author(s):  
Axel J. Giudicatti ◽  
Ariel H. Tomassi ◽  
Pablo A. Manavella ◽  
Agustin L. Arce

MicroRNAs are small regulatory RNAs involved in several processes in plants ranging from development and stress responses to defense against pathogens. In order to accomplish their molecular functions, miRNAs are methylated and loaded into one ARGONAUTE (AGO) protein, commonly known as AGO1, to stabilize and protect the molecule and to assemble a functional RNA-induced silencing complex (RISC). A specific machinery controls miRNA turnover to ensure the silencing release of targeted-genes in given circumstances. The trimming and tailing of miRNAs are fundamental modifications related to their turnover and, hence, to their action. In order to gain a better understanding of these modifications, we analyzed Arabidopsis thaliana small RNA sequencing data from a diversity of mutants, related to miRNA biogenesis, action, and turnover, and from different cellular fractions and immunoprecipitations. Besides confirming the effects of known players in these pathways, we found increased trimming and tailing in miRNA biogenesis mutants. More importantly, our analysis allowed us to reveal the importance of ARGONAUTE 1 (AGO1) loading, slicing activity, and cellular localization in trimming and tailing of miRNAs.


2020 ◽  
Author(s):  
Katarzyna Siudeja ◽  
Marius van den Beek ◽  
Nick Riddiford ◽  
Benjamin Boumard ◽  
Annabelle Wurmser ◽  
...  

AbstractTransposable elements (TEs) play a significant role in evolution by contributing to genetic variation through germline insertional activity. However, how TEs act in somatic cells and tissues is not well understood. Here, we address the prevalence of transposition in a somatic tissue, exploiting the Drosophila midgut as a model system. Using whole-genome sequencing of in vivo clonally expanded gut tissue, we map hundreds of high-confidence somatic TE integration sites genome-wide. We show that somatic retrotransposon insertions are associated with inactivation of the tumor suppressor Notch, likely contributing to neoplasia formation. Moreover, by applying Oxford Nanopore long-read sequencing technology, as well as by mapping germline TE activity, we provide evidence suggesting tissue-specific differences in retrotransposition. By comparing somatic TE insertional activity with transcriptomic and small RNA sequencing data, we demonstrate that transposon mobility cannot be simply predicted by whole tissue TE expression levels or by small RNA pathway activity. Finally, we reveal that somatic TE insertions in the adult fly intestine are found preferentially in genic regions and open, transcriptionally active chromatin. Together, our findings provide clear evidence of ongoing somatic transposition in Drosophila and delineate previously unknown underlying features of somatic TE mobility in vivo.


2020 ◽  
Vol 522 (3) ◽  
pp. 776-782
Author(s):  
Wei-Hao Lee ◽  
Kai-Pu Chen ◽  
Kai Wang ◽  
Hsuan-Cheng Huang ◽  
Hsueh-Fen Juan

2016 ◽  
Vol 13 (5) ◽  
Author(s):  
Matthew Kanke ◽  
Jeanette Baran-Gale ◽  
Jonathan Villanueva ◽  
Praveen Sethupathy

SummarySmall non-coding RNAs, in particular microRNAs, are critical for normal physiology and are candidate biomarkers, regulators, and therapeutic targets for a wide variety of diseases. There is an ever-growing interest in the comprehensive and accurate annotation of microRNAs across diverse cell types, conditions, species, and disease states. Highthroughput sequencing technology has emerged as the method of choice for profiling microRNAs. Specialized bioinformatic strategies are required to mine as much meaningful information as possible from the sequencing data to provide a comprehensive view of the microRNA landscape. Here we present miRquant 2.0, an expanded bioinformatics tool for accurate annotation and quantification of microRNAs and their isoforms (termed isomiRs) from small RNA-sequencing data. We anticipate that miRquant 2.0 will be useful for researchers interested not only in quantifying known microRNAs but also mining the rich well of additional information embedded in small RNA-sequencing data.


2009 ◽  
Vol 25 (18) ◽  
pp. 2298-2301 ◽  
Author(s):  
D. Langenberger ◽  
C. Bermudez-Santana ◽  
J. Hertel ◽  
S. Hoffmann ◽  
P. Khaitovich ◽  
...  

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