RNA Circular Dichroism Assay

In the field of molecular biology, understanding the intricate structures and functions of RNA molecules is crucial for unraveling the mysteries of life. Researchers rely on various techniques to analyze RNA, and one such powerful tool is Circular Dichroism (CD).

What is Circular Dichroism?

Circular Dichroism is a spectroscopic technique that measures the differential absorption of left- and right-circularly polarized light by optically active molecules. It provides valuable information about the secondary structure and conformational changes of biomolecules, including proteins, nucleic acids, and chiral small molecules.

What Can Circular Dichroism Provide for RNA Analysis

RNA molecules are essential players in gene expression, regulation, and numerous cellular processes. Understanding their secondary structure and folding dynamics is crucial for deciphering their functions. Circular Dichroism spectroscopy offers unique advantages in the analysis of RNA:

Secondary Structure Determination:

Circular Dichroism can probe the secondary structure of RNA molecules by assessing the differential absorption of circularly polarized light. The CD spectrum reflects the interactions between the RNA bases and their local environment, providing insights into the presence of helices, loops, and other structural motifs.

Folding and Conformational Changes:

The folding dynamics of RNA molecules play a crucial role in their biological activities. CD analysis allows researchers to monitor the folding kinetics and conformational changes of RNA in real-time. By measuring changes in the CD signal as a function of time, valuable insights can be obtained about the folding pathways, intermediate states, and stability of RNA molecules. This information is particularly relevant in studying RNA-protein interactions, ribozyme catalysis, and the development of RNA-based therapeutics.

Thermal Stability Analysis:

CD spectroscopy can be used to investigate the thermal stability of RNA structures. By monitoring changes in the CD signal as a function of temperature, researchers can generate melting curves that provide insights into the thermodynamic stability and folding cooperativity of RNA molecules. This information is valuable for determining the stability of RNA structures under different conditions and for comparing the stability of different RNA sequences or mutants.

The CD spectra of the duplexes (Gładysz et al., 2019)

Ligand Binding Studies:

Small molecules, proteins, and other nucleic acids frequently interact with RNA molecules. It is possible to use CD analysis to examine how RNA binds to its ligands. The binding affinity, stoichiometry, and structural alterations brought on by the interaction can all be determined by observing changes in the CD spectrum after ligand binding. This use is especially helpful in the drug development process since creating RNA-targeted therapies requires a thorough understanding of RNA-ligand interactions.

Comparative Studies:

The comparison of various RNA sequences, mutations, or variants is possible with RNA CD analysis. Researchers can spot variations in secondary structure, folding dynamics, or thermal stability between different RNA molecules by contrasting CD spectra or melting curves. These side-by-side analyses offer insightful information about the effects of sequence modifications or mutations on RNA structure and function.

Quality Control in RNA Research:

CD analysis can be utilized as a quality control tool for RNA samples. By assessing the CD spectra of RNA preparations, researchers can ensure the integrity and correct folding of RNA molecules. Any deviations or anomalies in the CD spectra can indicate potential issues with RNA sample quality, such as degradation or misfolding.

Creative Proteomics' RNA Circular Dichroism Analysis

Modern CD spectrometers provided by Creative Proteomics ensure exact measurements for trustworthy and reproducible data. Our staff of highly qualified biologists and spectroscopic specialists is well-versed in RNA analysis. On RNA molecules, we've successfully carried out a number of CD experiments that have given researchers everywhere useful information.

Recognizing the individuality of every RNA molecule, we collaborate closely with our clients to create assay methods that are specifically suited to their line of inquiry. Whether you need a CD assay for secondary structure research, folding kinetics analysis, or melting curve experiments, our experts will create one that is tailored to your requirements.

Furthermore, Creative Proteomics offers comprehensive data analysis services to complement the CD spectra. Our experts provide secondary structure prediction, thermal stability analysis, and folding kinetics modeling to help you interpret the results and extract meaningful information from your CD data.