Biotinylated Rna
Biotinylated Rna - Related Videos
11:32
Mapping RNA-RNA Interactions Globally Using Biotinylated Psoralen
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Cited by 20 •
2017
Here, we detail the method of Sequencing of Psoralen crosslinked, Ligated, and Selected Hybrids (SPLASH), which enables genome-wide mapping of intramolecular and intermolecular RNA-RNA interactions in vivo. SPLASH can be applied to study RNA interactomes of organisms including yeast, bacteria and humans.
07:55
An Optimized Quantitative Pull-Down Analysis of RNA-Binding Proteins Using Short Biotinylated RNA
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Cited by 3 •
2023
Here, we present an optimized in vitro method to uncover, quantify, and validate protein interactors of specific RNA sequences, using total protein extract from human cells, streptavidin beads coated with biotinylated RNA, and mass spectrometry analysis.
09:13
Cell-surface Biotinylation Assay
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2023
A cell can regulate the amount of particular proteins on its cell membrane through endocytosis, following which cell surface proteins are effectively sequestered in the cytoplasm. Once within a cell, these surface proteins can be either destroyed or “recycled” back to the membrane. The cell surface biotinylation assay provides researchers with a way to study these phenomena. The technique makes use of a derivative of the small molecule biotin, which can label surface proteins and then be...
10:34
Detection of RNA-binding Proteins by In Vitro RNA Pull-down in Adipocyte Culture
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Cited by 19 •
2016
An RNA pull-down protocol is optimized here for detection of interactions between RNA-binding proteins (RBPs) and noncoding as well as coding RNAs. An RNA fragment from androgen receptor (AR) was used as an example to demonstrate how to retrieve its RBP from lystate of primary brown adipocytes.
06:21
Depletion of Ribosomal RNA for Mosquito Gut Metagenomic RNA-seq
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Cited by 20 •
2013
A ribosomal RNA (rRNA) depletion protocol was developed to enrich messenger RNA (mRNA) for RNA-seq of the mosquito gut metatranscriptome. Sample specific rRNA probes, which were used to remove rRNA via subtraction, were created from the mosquito and its gut microbes. Performance of the protocol can result in the removal of approximately 90-99% of rRNA.
FAQs
The key mechanism is the selective recognition between the biotin attached to RNA and streptavidin or avidin. Because this interaction is strong, the tagged molecule can be retained on a coated surface or recovered with magnetic beads. This converts molecular recognition into a practical way to detect or isolate a specific RNA population.
Covalent attachment matters because it keeps the biotin associated with the same RNA molecule during downstream analysis. The tag provides a handle for immobilization or recovery, while the design preserves access to the nucleotide sequence and functional regions. Consequently, researchers can use the labeled RNA in assays that depend on sequence recognition or biological activity.
Coated surfaces and magnetic beads provide different formats for the same recognition principle. A surface supports immobilization for assays such as hybridization, whereas beads provide a recoverable material for isolation or pull-down experiments. The choice therefore depends on whether the main goal is an assay at a fixed interface or physical recovery of associated molecules.
Preserving RNA’s biological activity makes the tag useful beyond simple detection. A labeled transcript can still be examined for interactions with proteins or other nucleic acids, while biotin-streptavidin recognition supplies a way to retain or isolate the RNA. This combination connects molecular interaction studies with analysis of gene regulation.
A basic workflow starts with a biotinylated RNA preparation, exposes it to streptavidin- or avidin-compatible coated surfaces or magnetic beads, and then uses the retained RNA for the selected assay. Depending on the design, the result can support detection, purification, hybridization, localization, or interaction analysis.
For hybridization assays, the nucleotide sequence remains the informative feature, while the biotin tag provides a means to position or recover the RNA. In transcript localization studies, this pairing helps connect where a transcript is detected with the molecular identity of that RNA. Thus, labeling supports both spatial analysis and sequence-dependent readouts.
Pull-down experiments use the capture handle to examine molecules associated with the labeled RNA. Researchers can investigate RNA-protein or RNA-nucleic acid interactions by isolating the RNA-containing material and analyzing the interaction context. This makes the method useful for studying regulatory relationships, especially when the question concerns how RNA participates in gene regulation.