LncRNAs are over 200 nucleotides in length, lack a protein-coding potential, can be spliced, capped, and/or polyadenylated, and are localized either in the nucleus or the cytoplasm of the cell. Currently, it is difficult to obtain the tertiary structure of RNA, but there is partial evidence for the regulatory mechanism of lncRNAs through local single-stranded and local secondary structures. RNA primary structure is the nucleotide sequence consisting of bases AUGC in a specific order, and RNA secondary structure is the Watson-Crick double helix structure in which single-stranded RNA molecules fold to form G≡C and A=U base pairing, which also allows for G-U wobble base pairs.

Fig. 1 Differences in the structure and sequence between mRNA and lncRNA. (Li R et al., 2016)

Lifeasible provides a structural analysis of plant lncRNA by experimental methods, including enzymatic footprinting, chemical probing, nuclear magnetic resonance, and small angle scattering, to help our customers worldwide in plant science research. Our platform is equipped with cutting-edge facilities and professional experts to support research. Here, we provide various services according to customers' demands.

Enzymatic Foot-printing

• Enzymatic foot-printing is an in vitro approach designed to specifically cleave either single- or double-stranded nucleotides of radioisotope-labeled RNAs using ribonucleases (RNases) with different specificities. It can thus be used to map single- versus double-stranded regions of RNA.
• We provide structural analysis of lncRNA by enzymatic foot-printing, including parallel analysis of RNA structure and fragmentation sequencing. Then, the fragmented RNA products are typically analyzed alongside a sequencing or an alkaline hydrolysis RNA ladder by denaturing polyacrylamide gel electrophoresis.

Chemical Probing

• Chemical probing uses chemicals, which covalently modify atoms of single-stranded or flexible nucleotides. The high variety of chemicals (DMS, DEPC, CMCT) allows probing in vivo and in vitro. Reverse transcription introduces mutations into the cDNA, which subsequently can be mapped.
• We provide structural analysis of lncRNA by chemical probing, including SHAPE (selective 2′-hydroxyl acylation by primer extension) and PARIS (psoralen analysis of RNA interactions and structures). We also help our clients test SHAPE and PARIS reagents to improve the signal-to-background ratio and develop methodical improvements.

Nuclear Magnetic Resonance

Nuclear magnetic resonance (NMR) spectroscopy is a versatile biophysical and structural biology technique, thus representing a potent tool. We help our customers analyze the structure of lncRNA by NMR to determine precise base-pairing information and high-resolution three-dimensional RNA structures.

Small Angle Scattering

Small-angle scattering (SAS) is the collective name given to various techniques, including X-ray (SAXS) and neutron (SANS) scattering. We provide structural analysis of lncRNA by SAS to characterize lncRNA, such as the averaged particle sizes and shapes.

Lifeasible provides cost-effective, high-quality, and hassle-free services to our customers worldwide. We provide our clients with direct access to our experts and prompt responses to their questions. If you are interested in our services or have questions, please contact us or make an online inquiry.

Reference

1. Li R, et al. (2016). "Understanding the Functions of Long Non-Coding RNAs through Their Higher-Order Structures." Int J Mol Sci. 17 (5), 702.

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