Exosomes are small membrane vesicles composed of RNA, proteins, lipids, and bioactive metabolites. They are responsible for intercellular communication and take a part in immune systems. To analyze these data, a number of tools have been launched to be identify molecular groups. With the advent of Next generation sequencing (NGS) techniques, the research on exosomes is accelerated in various diseases including cancers. Currently, it is reported that bioinformatics-based analysis of RNA-Sequencing (RNA-Seq) data of exosomes from Trypanosoma cruzi has indicated the relevance of extracellular vesicles to many important gene products. Creative Biolabs is equipped with seasoned experts and advanced platforms that can assist global followers in high-efficient and high-quality RNA sequencing cases.
The exosome is a heterogeneous concept of bilayered lipid vesicle that originates from cytosolic compartments. This kind of vesicle ranges in diameter from 30nm to 150 nm. Usually, almost all cell types are known to be capable of secreting exosomes in physiological and pathological processes. The endocytic origin exosomes are thought to play essential roles in a variety of biological functions, especially cell-cell communications. Despite first described in 1983, the concentrations in exosomes have really strikingly increased after the discovery that they include mRNA and microRNA. This finding promotes renewed interest for the general field of secreted vesicles implicated in intercellular communications, and studies on their structures have exponentially grown over the last few years, exploring their compositions and functions, as well as their attractive value as biomarkers.
Fig.1 Exosomes biogenesis. (Zhou, 2017)
RNA-Seq refers to methods used to determine the specific sequence of RNA molecules. Unlike hybridization-based detection, it allows genome-wide analysis of transcription at a single nucleotide, including confirmation of alternative splicing and post-transcriptional RNA editing. RNA-Seq involves high-throughput shotgun sequencing of cDNA molecules harvested from RNA reverse transcription, and the NGS technique to sequence the RNA within a sample in an effort to identify the primary sequence and relative abundance of each RNA molecule.
Fig.2 Experimental design and sequencing platforms. (Li, 2014)
All RNA-seq workflows follow a similar protocol. Total RNA is extracted from a biological sample of interest which may be purified to enrich for mRNAs, microRNAs, lincRNAs, prior to preparing an RNA library. Sequencing can display one read in a single-end sequencing reaction, or two ends separated by an unsequenced fragment in paired-end reactions. In short, RNA-Seq has offered an unparalleled view of the transcriptome in both normal and pathological conditions. NGS is rapidly becoming an approach of choice for transcriptional profiling assays. Compared to microarray technology, high-throughput sequencing enables identification of novel transcripts, avoiding a sequenced genome and circumvents background noise linked to fluorescence quantification.
In recent years, exosomes have received tremendous attention because of biological fingerprints of exosomes practically reflect their parental cells. Although the exact function of exosomes remains to be fully dissected, cumulative evidence has demonstrated that exosomes play a vital role in varieties of cellular processes, including intercellular communication, antigen presentation, coagulation, waste management, and transfer of proteins and nucleic acids. As such, not only do exosomes contribute to a better understanding of cell pathophysiology, but have a great potential to be translated into many clinical practices, ranging from diagnosis and prognosis to nucleic acid delivery and disease therapeutics.
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.
In Creative Biolabs, we offer this high-throughput sequencing with numerous advantages over other gene expression experiments.
Creative Biolabs is skilled at providing custom exosome projects for basic research and clinical trials. If you don’t find exosome services you interest, please focus on these linkages or contact us with your particular needs.