Transfer RNA (tRNA) is a molecule that decodes mRNA and translates proteins. tRNA produces two degradation products through a precise biogenesis process: tiRNA (tRNA halves) and tRF (tRNA-derived fragments). In recent years, next-generation sequencing offers unprecedented opportunities to discover and quantify a wide variety of tRFs and tiRNAs. RNA-seq is an effective tool for high-throughput detection of tRF and tiRNA expression, and researchers has designed a variety of specific amplification primers to verify them in the database by qRT-PCR.
Fig 1. tRF & tiRNA-seq experiment workflow. (Fang Y, 2021)
In general, tRFs and tiRNAs exert their biological functions by regulating translation, gene expression, mRNA stability, epigenetics, and chromatin modification.
Fig 2. tRFs and tiRNAs regulate translation. (Xie Y, 2020)
BOC Sciences provides tRF&tiRNA sequencing services to offer insights into the study of biological functions and their underlying mechanisms. We are committed to helping our clients advance their next steps in RNA research. With the expertise in tRF&tiRNA, BOC Sciences tRFs&tiRNA sequencing service offers:
Quality control is performed using gel electrophoresis (GC) to identify the quantity and integrity of each RNA sample, followed by a spectrophotometer for total RNA quantification and quality assurance.
Pretreatment of total RNA samples to remove RNA modifications that may interfere with small RNA-seq library construction.
The sequencing library is constructed by using a commercial kit, which included 3' and 5' small RNA adapters, cDNA synthesis and library PCR amplification. Therefore, the influence of miRNA and other small RNA fragments can be excluded. At BOC Sciences, the prepared tRF and tiRNA-seq libraries are fully quantified.
Sequencing reactions are performed on based on the manufacturer's guide.
We evaluate tRF and tiRNAs levels using sequencing counts. tRF and tiRNA differentially expressed are screened based on calculated values using the R package EdgeR.
Depending on the miRNA target predictors, our experts use different algorithms to obtain the seed sequences and targets for tRFs and tiRNAs.
BOC Sciences provides gene annotation and analysis services, which can be used for pathway and process enrichment analysis of target genes.
We perform RT-qPCR verification for candidate tRFs and tiRNAs.
tRFs (tRNA-derived fragments) are typically 14-30 nucleotides from tRNA ends, while tiRNAs (tRNA halves) are 29-50 nucleotides produced by specific cleavage in the anticodon loop under cellular stress.
tRFs and tiRNAs regulate gene expression, translation, and mRNA stability, and play roles in cellular stress response, making them valuable for studying gene regulation mechanisms.
We use specialized library preparation for small RNAs, comprehensive reference databases, and advanced bioinformatics to precisely map cleavage sites and distinguish different fragment types.
Our service supports various sample types including cell lysates, tissue extracts, plasma, serum, and other biofluids, with optimized protocols for low-input and challenging samples.
We provide differential expression analysis, fragment classification, length distribution profiling, target prediction, and functional annotation to support comprehensive data interpretation.
Specific tRF/tiRNA expression patterns and fragment ratios can indicate biological states, making them valuable markers for cellular processes and comparative studies across experimental conditions.
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