What is exosomal RNA?

Exosomes are RNA and protein-containing small vesicles (30–150 nm) constantly secreted by all cells in culture and in vivo, in both a normal and disease state [1–3]. More importantly, exosomes deliver macromolecular messages (RNA and protein) that enable cell-to-cell communication and signalling [1,6–9].

What do extracellular vesicles do?

Extracellular vesicles (EVs) are lipid bilayer-delimited particles that are naturally released from almost all types of cell and, unlike a cell, cannot replicate. They carry a cargo of proteins, nucleic acids, lipids, metabolites, and even organelles from the parent cell.

How does small RNA Seq work?

Small RNA sequencing (Small RNA-Seq) is a type of RNA sequencing based on the use of NGS technologies that allows to isolate and get information about noncoding RNA molecules in order to evaluate and discover new forms of small RNA and to predict their possible functions.

What is RNA Seq used for?

RNA-seq (RNA-sequencing) is a technique that can examine the quantity and sequences of RNA in a sample using next-generation sequencing (NGS). It analyzes the transcriptome, indicating which of the genes encoded in our DNA are turned on or off and to what extent.

How do you isolate an exosome?

Various methods for the isolation of exosomes from biological fluids have been developed. They include centrifugation, chromatography, filtration, polymer-based precipitation and immunological separation. Recent technical improvements in these methods have made the isolation process faster and easier.

How do you isolate exosomes from serum?

The reagent is added to the serum sample, and the solution is incubated at 2°C to 8°C for 30 min. The precipitated exosomes are recovered by standard centrifugation at 10,000 x g for 10 min.

What is the main function of extracellular vesicles in unicellular and multicellular organisms?

All living organisms secrete molecules for intercellular communication. Recent research has revealed that extracellular vesicles (EVs) play an important role in inter-organismal cell-to-cell communication by transporting diverse messenger molecules, including RNA, DNA, lipids and proteins.

What is in extracellular vesicles?

Extracellular vesicles (EVs) are lipid bound vesicles secreted by cells into the extracellular space [1,2]. The content, or cargo, of EVs consists of lipids, nucleic acids, and proteins—specifically proteins associated with the plasma membrane, cytosol, and those involved in lipid metabolism [1,4].

What does ATAC seq measure?

The assay for transposase-accessible chromatin with sequencing (ATAC-Seq) is a popular method for determining chromatin accessibility across the genome. By sequencing regions of open chromatin, ATAC-Seq can help you uncover how chromatin packaging and other factors affect gene expression.

How does Chipseq work?

How Does ChIP-Seq Work? ChIP-Seq identifies the binding sites of DNA-associated proteins and can be used to map global binding sites for a given protein. ChIP-Seq typically starts with crosslinking of DNA-protein complexes. Samples are then fragmented and treated with an exonuclease to trim unbound oligonucleotides.

How does bulk RNA-Seq work?

Bulk RNA-Seq experiments provide a view of gene expression of an entire sample. This is done by dissociating the sample into individual single cells, identifying the cell types, and measuring the expression products of each cell.

How are exosomal RNA sequences prepared for RNA Seq?

However, detailed protocols describing exosome purification for preparation of exosomal RNA-sequence (RNA-Seq) libraries are lacking. Here we compared methods for isolation of exosomes and extraction of exosomal RNA from human cell-free serum, as well as strategies for attaining equal representation of samples within pooled RNA-Seq libraries.

Is it possible to extract exosomal RNA from Sera?

We determined the minimal starting volume of serum required for exosome preparation and showed that high quality exosomal RNA can be isolated from sera stored for over a decade. Finally, RNA-Seq libraries were successfully prepared with exosomal RNAs extracted from human cell-free serum, cataloguing both coding and non-coding exosomal transcripts.

How are messenger RNAs transported through the exosome?

Exosomal messenger RNAs (mRNAs) are transported via exosomes and can be translated into proteins once brought into recipient cells, indicating exosomal mRNAs play a regulatory role in non-cell autonomous signaling [ 22 ].

How are RNA-Seq and NGS used in medicine?

Today, RNA-Seq with NGS is an emerging method for scientists to study transcriptome. High-throughput NGS has been fastly adapted to various aspects of biomedical research and gradually engaged with certain clinical applications. Actually, NGS has already begun to produce visible benefits in some healthcare setting.