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Updated: Jul 7, 2026
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MISSION esiRNA for RNAi Screening in Mammalian Cells
Published on: May 12, 2010
ATP requirements and small interfering RNA structure in the RNA interference pathway
A Nykänen1, B Haley, P D Zamore
1Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, 55 Lake Avenue North, Worcester, MA 01655, USA.
Cell
|November 10, 2001
Summary
Adenosine triphosphate (ATP) drives key steps in RNA interference (RNAi), including siRNA processing and activation. This ensures only authentic small interfering RNAs (siRNAs) with 5' phosphates direct target RNA destruction.
Area of Science:
- Molecular Biology
- Biochemistry
- Gene Regulation
Background:
- RNA interference (RNAi) is a crucial gene silencing mechanism.
- The precise molecular steps and energy requirements of RNAi are not fully elucidated.
Purpose of the Study:
- To investigate the role of Adenosine triphosphate (ATP) in the RNA interference (RNAi) pathway.
- To delineate the sequential steps involved in RNAi-mediated gene silencing.
Main Methods:
- Biochemical assays to analyze RNA processing and complex formation.
- Enzyme kinetics to determine ATP dependency of specific steps.
- RNA cleavage assays to assess target recognition.
Main Results:
- Identified two critical ATP-dependent steps in the RNAi pathway.
- Demonstrated ATP-dependent processing of double-stranded RNA into small interfering RNAs (siRNAs).
- Revealed ATP-dependent unwinding of siRNA duplexes for activation and ATP's role in maintaining 5' phosphates on siRNAs.
Conclusions:
- RNAi involves at least four sequential steps, with key processing and activation stages requiring ATP.
- The 5' phosphate on siRNAs acts as a quality control mechanism, ensuring only authentic siRNAs mediate target RNA destruction.
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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...
01:23RNA 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...
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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...
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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.
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The pentose sugar in DNA is deoxyribose, while in RNA the pentose sugar is ribose. The difference between the sugars is the presence of the hydroxyl group on the ribose's second carbon and a hydrogen on the deoxyribose's second carbon. The phosphate residue attaches to the hydroxyl group of the 5′ carbon of one sugar and the hydroxyl group of the 3′ carbon of the sugar of the next nucleotide, which forms a 5′ to 3′ phosphodiester linkage.
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