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How do you inhibit mRNA degradation?

How do you inhibit mRNA degradation?

Degradation of both mRNAs was strongly inhibited in cells exposed to UV-B light. Removal of the poly(A)-tail, considered a crucial step in mRNA degradation, was strikingly impaired. UV light also inhibited deadenylation and degradation of endogenous mRNA of the chemoattractant cytokine interleukin (IL)-8.

What is mRNA degradation control?

mRNA degradation is controlled, at least in part, by microRNAs (miRNAs) and small-interfering RNAs (siRNAs). These small RNAs (~21–26 nucleotides in length) can trigger endonuclease cleavage or possibly they can stimulate the rate of mRNA decapping.

What increases mRNA degradation?

Several studies have shown that mutating the cap-binding complex to reduce initiation rates increases mRNA degradation. Reducing initiation rates using mutations in the cap-binding complex leads to enhanced mRNA decay by increasing the rates of deadenylation and decapping.

What inhibits mRNA translation?

Repression of translation by miRNAs can occur through inhibition of translation initiation or elongation, and may also lead to changes in the status of the mRNA 3′ poly(A) tail. Elements found within the mRNA 5′ UTR (yellow box) can bind regulatory proteins that repress translation by inhibiting 48S ribosome scanning.

What is the most common pathway for mRNA degradation?

mRNA degradation predominantly proceeds through two alternative routes: the 5′→3′ pathway, which requires deadenylation followed by decapping and 5′→3′ hydrolysis; and the 3′→5′ pathway, which involves deadenylation followed by 3′→5′ hydrolysis and finally decapping.

How is mRNA degradation detected?

In case you want to do detection of a specific mRNA in your cell lines I would recommend doing northern-blotting or reverse-transcription PCR. Maybe the easiest way is to do a quantitative PCR. If you see reduced levels of your mRNA, it could be interpreted as RNA degradation.

What determines mRNA degradation?

mRNA turnover is determined by the rates of mRNA synthesis and degradation, which jointly adjust the level of gene expression [1,2,3]. The degradation of mRNAs in eukaryotes occurs largely in the cytoplasm and begins with the removal of the poly(A) tail.

How is mRNA translation regulated?

Control of mRNA translation is mediated by a complex web of overlapping mechanisms, including, but not limited to, regulation of translation initiation and the length of the poly(A) tail, by the presence of short upstream open reading frames (uORFs), and by sequence-specific association with RBPs.

Why is RNA so easily degraded?

There are two main reasons for RNA degradation during RNA analysis. RNA is made up of ribose units, which have a highly reactive hydroxyl group on C2 that takes part in RNA-mediated enzymatic events. This makes RNA more chemically labile than DNA. RNA is also more prone to heat degradation than DNA.