Screening Service of RNA Interacting with Plant Proteins

CD BioSciences provides clients with plant protein interaction RNA screening services based on RNA-binding protein immunoprecipitation (RIP) technology. Our services to research the interaction between RNA and plant proteins can be used for analyzing gene expression, marker detection, and drug screening. It is also applied to research the interaction between viral RNA and plant proteins to reveal the mechanism of plant resistance to viral diseases.

What is RIP?

RIP technology utilizes antibodies against target proteins to precipitate the corresponding RNA-protein complexes. After isolation and purification, the RNAs on the complexes can be verified by q-PCR or sequencing analysis. RIP research of intracellular RNA-protein binding is a powerful way to understand the dynamics of post-transcriptional regulatory networks and help discover miRNAs' regulatory targets.

Figure 1. Schematic representation of RIP protocols. (Gagliardi, M., & Matarazzo, M. R., 2016)

Service Content

Project workflow

As an expert in protein-nucleic acid interaction research, we have established an efficient RIP technology platform to provide one-stop service. Our experimental process is as follows

• Cell or tissue samples are cross-linked to stabilize proteins and interacting RNA.
• Samples are lysed to release RNA and proteins into solution.
• Specific antibodies recognize and bind to the target protein, forming an immune complex.
• The immunocomplex is precipitated using protein A/G magnetic beads to separate unbound RNA and protein.
• The precipitated immune complex is washed several times to remove non-specific binding.
• The cross-links between RNA and protein are released by heat or enzyme treatment.
• Extract the RNA and perform subsequent analysis (RT-PCR, RNA sequencing, or microarray).

Problems we can solve

Notification for Sample Preparation and Handling

Information on the target protein.
Antibody to the target protein (available for IP) and its instructions.
Cell samples to be tested.
RNA ≥ 100ng (concentration ≥ 3ng/μL, OD260/280 ≥ 1.8, OD260/230 ≥ 1.5) to ensure that RNA is not significantly degraded.

Deliverables

1. Results pictures (including negative controls).
2. Data quality control.
3. Comparison with the reference genome.
4. Peak calling.
5. Sequence analysis.
6. Peak annotation.
7. Functional analysis of peak-related genes.
8. Visualization of RIP-seq data.

Application Fields

• Screens RNAs that interact with plant target proteins.
• Identify genome-wide networks of RNA-RBP (RNA binding protein) interactions.
• Analyze plant RBP interactions with non-coding RNAs (ncRNAs), long non-coding RNAs (lncRNAs), and micro RNAs (miRNAs).

CD BioSciences has rich experience in RNA-protein interaction research. Our professional technical team can provide clients with one-stop and efficient RIP services. Our competitive prices and extensive expertise have earned us the trust of our collaborators. Please contact us for more information.

For research use only, not for clinical use.