Rbp-rna Affinity
Rbp-rna Affinity - Related Videos
05:07
RNA-Protein Pull-Down Assay to Isolate RNA-Binding Proteins via Affinity Extraction
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2025
This video demonstrates an in vitro RNA pull-down assay to identify RNA-binding proteins (RBPs), which interact with the adenylate-uridylate-rich element (ARE) sequences in mRNA. The target RBPs from a cell lysate are mixed with an RNA probe to form RNA-protein complexes. The complexes are isolated via affinity purification utilizing the affinity of the desthiobiotin label of the RNA probe to a streptavidin-labeled magnetic bead.
02:30
Isolation of Small Regulatory RNA–Bound Bacterial Target RNA Using Affinity Purification
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2025
Source: Mercier, N., et. al., MS2-Affinity Purification Coupled with RNA Sequencing in Gram-Positive Bacteria. J. Vis. Exp. (2021)This video demonstrates the isolation of MS2-tagged small regulatory RNA (sRNA) bound to its bacterial target RNA using affinity purification with a maltose-binding protein (MBP)–MS2 coat protein fusion immobilized on amylose resin.
03:07
Electron Affinity
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2020
The electron affinity (EA) is the energy change for adding an electron to a gaseous atom to form an anion (negative ion). This process can be either endothermic or exothermic, depending on the element. Many of these elements have negative values of EA, which means that energy is released when the gaseous atom accepts an electron. However, for some elements, energy is required for the atom to become negatively charged, and the value of their EA is positive. Just as with ionization energy,...
01:41
Affinity and Avidity
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2019
Overview Antibodies bind to toxins or substances on the surface of cells, bacteria, viruses, or fungi. The substance is called an antigen, and the precise binding site is the epitope. The strength of the antibody-epitope interaction is called affinity. When an antibody binds an antigen by multiple epitopes, the cumulative strength of the interaction is called avidity. The strength of the interaction influences the elicited immune response. The Adaptive Immune System Increases Efficiency by...
08:34
MS2-Affinity Purification Coupled with RNA Sequencing in Gram-Positive Bacteria
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Cited by 10 •
2021
MAPS technology has been developed to scrutinize the targetome of a specific regulatory RNA in vivo. The sRNA of interest is tagged with a MS2 aptamer enabling the co-purification of its RNA partners and their identification by RNA sequencing. This modified protocol is particularly suited for Gram-positive...
FAQs
RNA sequence provides recognizable features, while secondary structure determines whether those features are exposed or constrained within the molecule. An RBP can therefore respond to both the nucleotide arrangement and the RNA’s structural configuration. Changes in either feature may alter molecular recognition, helping explain why related RNA molecules can exhibit different binding behavior under the same conditions.
Protein-binding domains contribute defined molecular surfaces that contact RNA, while electrostatic interactions help support association between the protein and the nucleic acid. Their combined effects influence both the strength and selectivity of binding. Examining these contributions can clarify how alterations in an RBP change recognition of particular RNA elements involved in gene regulation.
Affinity analysis can quantify how readily an RBP associates with an RNA molecule and how readily the complex dissociates when conditions change. Measuring these behaviors under defined conditions provides a basis for comparing interactions among different RNA or protein variants. The resulting comparisons reveal whether a molecular change affects binding strength, recognition, or both.
Measurements should use defined experimental conditions so that differences in association or dissociation can be attributed to the RNA, the RBP, or their interaction rather than uncontrolled variation. Maintaining consistent conditions is especially important when comparing sequence changes, structural differences, protein domains, or electrostatic contributions across multiple binding analyses.
A typical analysis begins by selecting the RBP and RNA elements of interest, then examining their interaction under defined conditions. Researchers measure association or dissociation and compare the results across relevant RNA or protein forms. This workflow can identify regulatory RNA elements and show how changes in either binding partner modify molecular recognition.
In genetics, these measurements help connect molecular recognition with post-transcriptional regulation, including splicing, RNA stability, localization, translation, and degradation. They are also useful for examining genetic variation and disease-associated mutations. By comparing altered and unaltered RNA or proteins, researchers can assess whether a change may influence regulatory interactions.