Translatome Profiling
Translatome Profiling - Related Videos
09:13
In vivo Interrogation of Central Nervous System Translatome by Polyribosome Fractionation
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Cited by 14 •
2014
This protocol illustrates essential modifications of polyribosome fractionation in order to study the translatome of in vivo CNS samples. It allows global assessment of translation and transcription regulation through the isolation and comparison of total RNA to ribosome bound RNA fractions.
10:56
Polysome Fractionation and Analysis of Mammalian Translatomes on a Genome-wide Scale
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Cited by 217 •
2014
Ribosomes play a central role in protein synthesis. Polyribosome (polysome) fractionation by sucrose density gradient centrifugation allows direct determination of translation efficiencies of individual mRNAs on a genome-wide scale. In addition, this method can be used for biochemical analysis of ribosome- and polysome-associated factors such as chaperones and signaling molecules.
08:29
Expression Profiling with Microarrays
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2023
Microarrays are important tools for profiling gene expression, and are based on complementary binding between probes that are attached to glass chips and nucleic acids derived from samples. Using these arrays, scientists can simultaneously evaluate the expression of thousands of genes. In addition, the expression profiles of different cells or tissue types can be compared, allowing researchers to deduce how the expression of different genes change during biological processes, and thus gain...
07:13
Using Topographic Maps to Generate Topographic Profiles
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2023
Source: Laboratory of Alan Lester - University of Colorado Boulder Topographic maps are "plan-view" representations of Earth's three-dimensional surface. They are a standard type of map-view that provides an overhead, or aerial, perspective. Among the defining features of a topographic map are the contour lines that indicate locations of constant elevation. The elevation interval between the contour lines is dependent on the level of detail provided by the map and the kind of topography present.
09:16
Eukaryotic Polyribosome Profile Analysis
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Cited by 51 •
2010
This article describes a protocol for the extraction of translating ribosomes from eukaryotic cells. Once extracted, ribosomes are separated into monosomes and polyribosomes by sucrose gradient fractionation to allow different ribosomal populations to be analyzed. As such, this method is the gold standard for examining the regulation of translation.
FAQs
Translatome profiling adds a protein-production layer to transcriptional analysis by showing which messenger RNAs are being used for translation. A transcript may be present without equivalent protein output, so measuring active translation can expose regulatory changes that RNA abundance alone would miss. This distinction is useful when interpreting altered gene expression in neural tissue.
Ribosome occupancy is inferred from the locations of sequenced, protected mRNA fragments along their source transcripts. When those positions are mapped, they indicate where translating ribosomes were situated and can help distinguish transcripts engaged in protein production from RNAs that are not actively being translated. This positional information adds detail about translation across individual messenger RNAs.
Translational regulation can connect RNA use with changes in synaptic function, plasticity, and cellular stress. These processes depend on how neural cells adjust protein production in response to their state, activity, or disease-related conditions. Profiling this regulatory layer therefore helps explain how molecular changes in neurons may contribute to altered neural behavior or pathology.
A basic ribosome-profiling workflow isolates translating ribosomes, preserves the messenger RNA fragments protected by those ribosomes, sequences the fragments, and maps their positions to transcripts. The resulting distribution of protected sequences indicates which RNAs are engaged in translation and where ribosomes occupy them, providing a direct basis for analyzing protein-production activity.
Researchers can compare translation across brain regions, cell types, developmental stages, and neuronal states. They can also examine responses associated with neural activity or disease. These comparisons help identify whether protein-production patterns vary with cellular identity, maturation, physiological state, or pathology, giving translational changes a defined anatomical or biological context.
In neuroscience, translatome data can reveal how changes in RNA use relate to synaptic function, plasticity, cellular stress, and disease-associated states. By examining actively translated messenger RNAs rather than transcriptional changes alone, researchers can investigate protein-synthesis programs that may help explain differences in neural behavior and pathological processes.