Endogenous Mrna Visualization
Endogenous Mrna Visualization - Related Videos
07:00
Visualization and Analysis of mRNA Molecules Using Fluorescence In Situ Hybridization in Saccharomyces cerevisiae
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Cited by 25 •
2013
This protocol describes an experimental procedure for performing Fluorescence in situ Hybridization (FISH) for counting mRNAs in single cells at single-molecule resolution.
08:33
Cell-Free In Vitro mRNA Translation In Drosophila Lysates
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2026
Here, we provide details on how to prepare lysates and perform in vitro translation using Drosophila embryos.
02:22
Regulated mRNA Transport
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2020
In eukaryotes, transcription and translation are compartmentalized; an mRNA is first synthesized in the nucleus and then selectively transported to the cytoplasm for protein synthesis. Before transport, a pre-mRNA undergoes several steps of post-transcriptional modifications including splicing, 5' capping, and the addition of a poly-adenine tail. Various proteins bind to the pre-mRNA during these modifications. The mRNA transport takes place with the help of multiple proteins playing specific...
02:31
Nuclear Export of mRNA
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2020
Before mRNAs are exported to the cytoplasm, it is crucial to check each mRNA for structural and functional integrity. Eukaryotic cells use several different mechanisms, collectively known as mRNA surveillance, to look for irregularities in mRNAs. Irregular or aberrant mRNA are rapidly degraded by various enzymes. If a defective mRNA escapes the surveillance, it would be translated into a protein which would either be non-functional or not function properly. One of the primary irregularities in...
02:01
pre-mRNA Processing
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2019
In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA. Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a “cap” to the 5’ end of the growing transcript. In this process, a 5’ phosphate is replaced by modified guanosine that has a methyl group attached to it (7-Methyl guanosine). This 5’ cap helps the...
FAQs
Detection depends on nucleic acid hybridization: a fluorescent probe binds a complementary region of the target transcript. Bound probes generate signals whose abundance and subcellular distribution can be examined, linking where transcripts occur with how strongly they are represented in the sampled cell or tissue. This preserves native gene-expression context.
Artificial overexpression can change the amount of transcript present, whereas measuring naturally produced RNA keeps the observation tied to the cell's existing biological state. That distinction is important when interpreting transcript abundance or location, because the resulting pattern reflects native production and distribution rather than an experimentally increased RNA signal.
Fixed-sample imaging and live-cell adaptations answer different biological questions. Fixed samples provide a preserved snapshot of transcript distribution, while live-cell formats can be adapted to monitor RNA localization and movement over time. Choosing between them therefore depends on whether the study prioritizes spatial mapping at one state or dynamic behavior in living cells.
A practical workflow begins by selecting the transcript of interest and using fluorescent probes designed to bind complementary sequence regions. The probes are then applied to either fixed samples or a format adapted for living cells, followed by imaging. Researchers interpret the resulting signal in relation to transcript abundance and subcellular distribution.
The resulting images provide more than a simple presence-or-absence readout. They can show how much target RNA is represented and where it is distributed within a cell or tissue. Those measurements help connect transcript patterns with cellular function and support investigation of transcription, RNA transport, translation, cell differentiation, and development.
By preserving information about transcript location, the method helps researchers relate gene-expression patterns to cellular organization and function. In biology, that perspective is relevant to processes such as differentiation and development, as well as transcription, transport, and translation. In disease research, transcript distribution or abundance can be examined within native cells or tissues.