Introduction of lncRNA
lncRNA: lncRNA is an abbreviation for long non-coding RNA molecule between 200 and 100,000 nt in length. Most lncRNA have significant spatiotemporal expression specificity during tissue differentiation and development. The sequence conservation of lncRNA is low, and only about 12% of lncRNA can be found in organisms other than humans.
According to the position of lncRNA in genome relative to protein-coding genes, they can be divided into 7 categories: promoter-associated, intronic, bidirectional, sense, antisense, 3’ UTR associated, intergenic. Moreover, the positional relationship is a tool to speculate the function of lncRNA.
Fig.1 Classification of lncRNA. (Huang X, 2018)
In general, lncRNA regulates gene expression in three aspects:
- Epigenetic regulation: lncRNA recruits chromatin remodeling complexes to specific sites and mediate silencing of related gene expression.
- Transcriptional regulation: including the following: A. lncRNA transcription can interfere with the expression of adjacent genes. B. lncRNA can interfere with gene expression by blocking promoter regions. C. lncRNA can interact with RNA binding proteins and localize them to gene promoter regions to regulate gene expression. D. lncRNA can regulate the activity of transcription factors. E. lncRNA can also regulate gene expression by regulating basic transcription factors.
- Post-transcriptional regulation: lncRNA can regulate gene expression by forming a double-stranded form with mRNA at the post-transcriptional level.
Generally, lncRNA's abundance in cells is a dozen times lower than mRNA. When lncRNA perform a regulatory function, they form a tight complex with DNA, RNA or protein molecules, and are selectively distributed in the nucleus and cytoplasm.
Coupled with the sensitivity and detection accuracy limitations, the traditional siRNA tools for mRNA functional deletion studies are likely to be unsuitable for lncRNA functional deletion studies due to their complexity.
The BOC Sciences' lncRNA expert team can provide you with a full range of services from lncRNA screening and validation, lncRNA functional exploration, deeper target search, and expression and regulation mechanism exploration.
- Bioinformatics prediction: Predict possible lncRNA based on transcriptome information and gene characteristics provided by comparing with database, as well as removing known possible protein-coding transcripts, tRNA, rRNA, repeat, etc.
- Sequencing: Reverse transcription of lncRNA into cDNA by reverse transcription method to determine the sequence of a specific segment, and apply the RACE method to determine the sequence at both ends.
- Bioinformatics analysis: screening of differential lncRNA and mRNA, cluster analysis, functional enrichment analysis, lncRNA-mRNA/microRNA correlation analysis, lncRNA target gene prediction.
- lncRNA overexpression: The lncRNA sequence is constructed into a viral vector, which can be used for transient transfection of cells or packaged into a virus for stable strain screening and overexpression of lncRNA at the animal level.
- lncRNA interference: The interference fragment is designed for lncRNA and constructed into a viral vector. The vector can be transiently transfected into cells for lncRNA interference or packaged as a lentivirus for lncRNA interference at the animal level.
- lncRNA knockdown: lncRNA is knocked down by CRISPR/Cas9 technology. Multiple sgRNA sequences are designed for each end of a specific fragment of lncRNA for large fragment deletion, thus achieving the knockdown effect. The sgRNA sequences are constructed into a viral vector, then Cas9 sequences are loaded into another vector. Subsequently, both viruses infect the target cells simultaneously for the knockdown.
- lncRNA target gene search: method similar to the miRNA dual luciferase assay indicated above.
- Functional study of lncRNA.
- BOC Sciences recruited a professional research team of lncRNA experts with excellent technical skills, focusing on comprehensive lncRNA service to provide you with an integrated lncRNA research solution.
- BOC Sciences has successfully completed numerous lncRNA projects and has proven lncRNA-related experience.
- If you need detailed guidance on lncRNA topics/experiments, please contact us to learn more with BOC Sciences' lncRNA experts.
- Huang X; et al. Advances in esophageal cancer: A new perspective on pathogenesis associated with long non-coding RNAs. Cancer Letters. 2018, 413: 94-101.