Target Mrna Cleavage
Target Mrna Cleavage - Related Videos
09:16
Analysis of RNA Processing Reactions Using Cell Free Systems: 3' End Cleavage of Pre-mRNA Substrates in vitro
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Cited by 1 •
2014
RNA polymerase II synthesizes a precursor RNA that extends beyond the 3' end of the mature mRNA. The end of the mature RNA is generated cotranscriptionally, at a site dictated by RNA sequences, via the endonuclease activity of the cleavage complex. Here, we detail the method to study cleavage reactions in vitro.
07:09
Using Cleavage Under Targets and Tagmentation (CUT&Tag) Assay in Mouse Myoblast Research
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2024
Researchers new to the epigenetic field will find CUT&Tag a significantly easier alternative to ChIP assays. CUT&Tag has tremendously benefited the epigenetic studies on rare and primary cell populations, generating high-quality data from very few cells. This protocol describes performing H3K4me1 CUT&Tag assays on mouse myoblasts isolated from mouse hindlimb muscles.
02:51
mRNA Stability and Gene Expression
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2020
The structure and stability of mRNA molecules regulates gene expression, as mRNAs are a key step in the pathway from gene to protein. In eukaryotes, the half-life of mRNA varies from a few minutes up to several days. mRNA stability is essential in growth and development. The absence of the proteins regulating its stability, such as tristetraprolin in mice, can cause systemic issues, including bone marrow overgrowth, inflammation, and autoimmunity. Cis-acting Elements involved in mRNA stability
09:48
Profiling of H3K4me3 Modification in Plants using Cleavage under Targets and Tagmentation
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Cited by 1 •
2022
Cleavage under targets and tagmentation (CUT&Tag) is an efficient chromatin epigenomic profiling strategy. This protocol presents a refined CUT&Tag strategy for the profiling of histone modifications in plants.
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...
FAQs
Sequence complementarity allows the small interfering RNA to recognize and pair with a matching region of the target messenger RNA. This pairing positions the RNA within an Argonaute-containing complex and identifies where cleavage should occur. Because recognition depends on the paired sequence, the process can be directed toward a selected transcript rather than producing a general reduction in cellular messenger RNA.
The catalytic Argonaute component performs the cutting reaction after the small interfering RNA has guided the complex to its complementary messenger RNA sequence. Its activity converts sequence recognition into physical transcript cleavage. That cleavage is important because the affected messenger RNA becomes destabilized, linking molecular targeting to reduced availability of the transcript for protein production.
The process acts on messenger RNA after the transcript has been produced, rather than directly preventing transcription of the corresponding gene. Cleaving and destabilizing that RNA reduces the amount of message available for production of its encoded protein. This distinction helps separate effects on RNA regulation from mechanisms that act earlier during gene expression.
Reducing a selected messenger RNA provides a way to examine the consequences of lowering its encoded protein. Researchers can use this change to test whether the targeted gene contributes to a biological function or regulatory pathway. The approach therefore connects a defined RNA-level intervention with functional investigation, while avoiding the need to describe the gene's role only from its sequence.
Researchers direct a small interfering RNA toward the messenger RNA they want to suppress, allowing the complementary sequence and Argonaute-containing complex to act on that transcript. The resulting cleavage and destabilization reduce production of the associated protein. This logic supports selective reduction of unwanted gene expression and provides a focused way to investigate its biological consequences.
Targeted messenger RNA reduction can be used to perturb one component of a regulatory pathway and examine the effects of lowering its encoded protein. Because the intervention is directed to a chosen transcript through sequence complementarity, it helps researchers connect that component with pathway behavior. These studies can clarify gene relationships and identify consequences of altered expression.