Ribosome Profiling Sequencing (Ribo-seq) refers to the sequencing of RNA fragments bound to ribosomes that are being translated, to accurately obtain information and quantify all translatable molecules (including mRNA and other potentially translatable RNA molecules such as lncRNA, circRNA, etc.) in the sample, and to bridge the gap between the transcriptome and the proteome. Ribo-seq is the most commonly used translation sequencing technology, which utilizes RNA enzymes to digest RNA in cells to obtain ribosome-protected RNA footprints (RFs) that are being translated, and then enrich and deeply sequence these ribosome-protected RNA fragments of 30 nt or so, Ribo-seq can study the level, region, and rate of intracellular gene translation, which can be combined with transcriptome, small RNA sequencing, proteome and other correlation analyses to more accurately study the mechanisms of post-transcriptional and translational regulation.

Lifeasible has rich experience in the field of plant multi-omics, relying on our advanced technology platform and experienced technical team, we provide translational group sequencing that can help researchers with comprehensive and in-depth information on plant genome and transcriptome, which is of great significance in the field of crop improvement, biotechnology and so on.

Features of Ribo-seq technology

Ribo-seq can reveal when, where, and how proteins are being synthesized, which proteins are being synthesized, and the regulatory mechanisms of protein synthesis.Ribo-seq makes it possible to quantitatively translate a single nucleotide resolution on a genome-wide scale. Ribo-seq also has the advantage of being instantaneous, quantitative and localized compared to traditional means of studying protein translation. Ribo-seq has been widely used to study protein translation under different experimental and physiological conditions. Our Ribo-seq service bridges the gap from transcriptomics to proteomics, and has been widely used in plant research to reveal the regulatory mechanisms of growth and development, morphogenesis, disease, and stress.

Experimental procedure of Ribo-seq technology

• Plant sample collection and pretreatment. Collect target plant tissues, such as leaves, roots, etc., and freeze them quickly. Whole cell lysate is prepared by grinding the sample in liquid nitrogen.
• Ribosome protection step. The lysate was incubated with ribosome inhibitor to stabilize and protect the translating mRNA-ribosome complex. Ribosome-protected mRNA fragments were isolated by differential centrifugation.
• Extraction of mRNA fragments and sequencing library construction. Ribosome-protected mRNA fragments were extracted, and free non-protected mRNAs were removed, and sequencing libraries were constructed, including end repairs and junctions.
• High-throughput sequencing and data analysis. The constructed sequencing library was subjected to high-throughput sequencing to obtain Ribo-seq data. Bioinformatics tools were used to process and analyze the sequencing data.
• Interpretation and application of results. By analyzing the Ribo-seq data, we can understand the regulation of plant transcripts at the translation level, including translation efficiency, translation start site and other information.

Applications of Ribo-seq

• Adverse stress research. Environmental stress can limit the growth and development of plants and animals, and the study of translational regulation under stress conditions is of great value for resistance breeding. Currently, Ribo-seq has been widely used in the study of translational regulatory networks under plant adversity stresses, such as heat stress, light stress, drought stress, and oxidative stress.
• Tissue morphology study. Transform sequencing is of great significance in tissue morphology research, which can help researchers gain insight into the changes in gene expression regulation and protein synthesis levels in different tissue morphologies and reveal the mechanisms of plant and animal adaptations to the environment by revealing information on translational regulation, transcriptional start sites, and splicing variants, as well as mutations and polymorphisms.
• Growth and development. In higher plants, translational regulation plays an important role in different developmental processes, and it controls the expression of developmental, stage-specific, and tissue-specific gene products.
• Disease resistance mechanisms. Translome sequencing is also indispensable in the study of plant disease resistance mechanisms. When plants are invaded by pathogenic microorganisms, pattern-triggered immunity (PTI) and effector-triggered immunity (ETI) are triggered, and the induction of both types of immunity involves the activation of translation of a series of defense proteins; therefore, translational regulation plays a crucial role in the induction of plant immunity.

Plant tissue sample requirements

Why choose us?

• Experienced. So far, we have successfully completed Arabidopsis thaliana, poplar, tobacco, tomato, chrysanthemum, corn, soybean, rice, cotton and other tissues, etc., with rich service experience.
• Fast cycle time, good quality and high stability. We provide Ribo-seq using the advanced Illumina platform for sequencing, and combine it with High performance computing platform to achieve fast and stable sequencing data analysis and delivery. Lead time is <65 days.
• Professional protocol design and strict quality control. For plant tissues, we design different project programs and adjust test protocols, each step requires scientific and meticulous design to ensure high-quality research results.
• Professional bioinformatics analysis. We have a strong bioinformatics team to meet the needs of our clients for in-depth data analysis.

Lifeasible is committed to high-throughput technology in plant research and analysis from transcriptomics to proteomics, our advanced sequencing platform and experienced team of experts aim to provide plant researchers with international advanced high-throughput detection system solutions. If you are interested in us, please feel free to contact us.

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