Small RNA is a concept used to characterize many non-coding ribonucleic acid groups (ncRNA). While the term bacterial small RNA is used to define the short ncRNA chains present in bacterial cells, other types of ncRNA that have been examined in organisms, plants, and fungi also apply as an umbrella term. In gene regulation, small RNA is highly involved and may have used in gene therapy. In function, ncRNA varies from other RNAs since it does not code for a protein directly. Instead, ncRNA, like gene expression and modification, is included in other cellular functions. There are many types of small RNA, including microRNAs, short interfering RNAs (siRNAs), and tiny nucleolar RNAs (snoRNAs). In gene silencing, specific types of small RNA are essential, such as microRNA and siRNA. In chemical modifications of other RNA molecules, snoRNAs, another type of small RNA, function as mediators.
Small RNA Sequencing
In gene silencing and post-transcriptional regulation of gene expression, small noncoding RNAs operate. Small RNA sequencing is a tool to separate small RNA species and sequence them. Thousands of small RNA sequences with unparalleled sensitivity and dynamic range can be queried by small RNA-Seq.
With small RNA-Seq, novel miRNAs and other small non-coding RNAs can be discovered and the differential expression of these small RNAs in any sample can be studied. With single-base resolution, you can identify variations such as isomiRs (miRNA sequences that have variations with respect to the reference sequence), as well as evaluate any small RNA without previous sequence or secondary structure details.
The necessary first step in studies investigating post-transcriptional modulation of gene expression in health or disease is to recognize miRNA signatures in animal tissues. Small RNA sequencing is a next-generation sequencing-based transcriptomics method that is currently known as the most efficient and flexible miRNA profiling technique. The most commonly accepted NGS technology is Illumina synthesis sequencing (SBS) chemistry, which produces nearly 90% of sequencing data worldwide. Illumina provides an easy workflow for small RNA and miRNA sequencing, in addition to our industry-leading data consistency, which simplifies the whole procedure, from library planning to data processing and biological interpretation.
Small RNA sequencing has a lot of advantages over other methods in terms of unveiling the function of non-coding RNA. These include (1) understanding the influence of post-transcriptional regulation on phenotype, (2) identification of novel biomarkers, and (3) capturing the full range of species of small RNA especially miRNA.
Small RNA Sequencing in miRNA Profiling
miRNAs are short non-coding RNA molecules that, by insufficient complementarity to aim sequences typically found in the 30 UTR of mRNAs, downregulate gene expression. By reducing mRNA stability and/or by blocking translation, miRNAs act to decrease levels of corresponding target gene proteins.
Small RNA sequencing is an NGS technology that requires cDNA sequencing synthesized from a tissue sample's miRNA fragment after incorporation of adapters on both 30 and 50 ends and PCR amplification. The resulting raw data comprises specific reads later linked to sequences accessible on miRbase or to the annotated genome of mature miRNAs. Estimation of novel miRNAs is now possible because of the versatility of the post-sequencing review, and final data can be modified by remapping raw reads to the newest miRbase annotation.
References:
- Pantaleão LC, Ozanne SE. Small RNA Sequencing: A Technique for miRNA Profiling. InInvestigations of Early Nutrition Effects on Long-Term Health 2018 (pp. 321-330)..
