Abstract
We report the development of photo-crosslinked siRNA strands modified at each terminus with p-cyanostilbene. The siRNA was nuclease resistant and retained RNAi activity. We further studied the activation mechanism of the covalently-crosslinked siRNA. Interestingly Dicer, which is known to generate siRNA with overhanging 3' ends from the precursor siRNA, did not cleave the crosslinked siRNA at all. Our results suggest that the activation of the crosslinked siRNAs required cleavage by Argonaute2.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Argonaute Proteins / chemistry*
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Argonaute Proteins / metabolism
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Base Sequence
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Humans
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Oligonucleotides / chemistry*
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Oligonucleotides / metabolism
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Photochemical Processes
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RNA Interference / physiology
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RNA Interference / radiation effects*
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RNA, Small Interfering / genetics
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RNA, Small Interfering / metabolism
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RNA-Induced Silencing Complex / chemistry
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RNA-Induced Silencing Complex / genetics*
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RNA-Induced Silencing Complex / metabolism
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Stilbenes / chemistry*
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Stilbenes / metabolism
Substances
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Argonaute Proteins
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Oligonucleotides
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RNA, Small Interfering
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RNA-Induced Silencing Complex
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Stilbenes
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p-cyanostilbene