Related Experiment Video
Updated: Aug 12, 2026
08:55
In vitro Transcription and Capping of Gaussia Luciferase mRNA Followed by HeLa Cell Transfection
Published on: March 26, 2012
Sequential synthesis of small capped RNA transcripts in vitro by vesicular stomatitis virus
Virology
|February 1, 1983
Summary
Researchers identified small RNA species synthesized in vitro by vesicular stomatitis virus (VSV). These findings support a single entry model for VSV messenger RNA (mRNA) transcription.
Area of Science:
- Virology
- Molecular Biology
- RNA Synthesis
Background:
- Vesicular stomatitis virus (VSV) transcription involves complex mechanisms for messenger RNA (mRNA) synthesis.
- Previous models proposed simultaneous initiation of multiple transcripts at the onset of transcription.
Purpose of the Study:
- To investigate the mechanism of VSV mRNA transcription.
- To identify and characterize small RNA species synthesized in vitro.
- To provide evidence supporting or refuting existing transcription models.
Main Methods:
- In vitro transcription using purified VSV ribonucleoprotein (RNP) cores.
- Analysis of synthesized small RNA species (12-47 nucleotides).
- Characterization of RNA termini (capped vs. 5' triphosphate) and template properties.
Main Results:
- Identification of small capped RNA species originating from the N and NS genes.
- These capped RNAs were synthesized concomitantly with their respective mRNAs.
- Template analysis indicated specific initiation sites and sequential RNA appearance.
Conclusions:
- The findings challenge the multiple entry, start-stop model of VSV transcription.
- Evidence supports a single entry model for VSV mRNA synthesis.
- The study elucidates the coordinated synthesis of mRNA termini and full-length transcripts.
Related Concept Videos
01:23RNA Interference
RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
01:10Retrovirus Life Cycles
Retroviruses have a single-stranded RNA genome that undergoes a special form of replication. Once the retrovirus has entered the host cell, an enzyme called reverse transcriptase synthesizes double-stranded DNA from the retroviral RNA genome. This DNA copy of the genome is then integrated into the host’s genome inside the nucleus via an enzyme called integrase. Consequently, the retroviral genome is transcribed into RNA whenever the host’s genome is transcribed, allowing the retrovirus to...
02:28Leaky Scanning
During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA. Marilyn Kozak discovered that the sequence RCCAUGG (where R stands for...
02:30siRNA - Small Interfering RNAs
Small interfering RNAs, or siRNAs, are short regulatory RNA molecules that can silence genes post-transcriptionally, as well as the transcriptional level in some cases. siRNAs are important for protecting cells against viral infections and silencing transposable genetic elements.
In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the ATP-dependent...
In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the ATP-dependent...
01:26Size and Structure of Viral Genomes
Viral genomes exhibit remarkable diversity in size, structure, and composition, influencing their replication strategies and interactions with host cells. These genomes consist of either DNA or RNA and may be linear or circular. Additionally, they can be single-stranded or double-stranded, with each configuration affecting how the virus propagates within a host. RNA viruses, for instance, generally have smaller genomes than DNA viruses, a factor that contributes to their high mutation rates and...
01:29Viruses with RNA Genomes
RNA viruses are categorized into positive-strand, negative-strand, or double-stranded groups based on their genomic structure and replication mechanisms. This classification dictates how they exploit host cellular machinery for protein synthesis and replication. Some RNA viruses also utilize reverse transcription as part of their life cycle, further diversifying their replication strategies.Positive-Strand RNA VirusesPositive-strand RNA viruses have genomes that function directly as messenger...