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miRNA specificity controls: paper

miRNA specificity controls: paper - RNAi and SiRNA Forum

miRNA specificity controls: paper - Discuss and Post Questions in this forum about RNA interference and siRNA. RNAi and siRNA transfection, inhibition of endogenous genes, and miRNA and shRNA.


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Old 07-24-2007, 02:23 PM
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Default miRNA specificity controls: paper



Knockdowns of miRNA activity have generally used oligos targeting the miRNA guide strand. A Morpholino oligo targeting an miRNA guide strand can interfere with the activity of the miRNA. It is difficult to control for the specificity of the knockdown when using this technique alone. However, Morpholinos targeting the nucleolytic processing sites of an immature miRNA can prevent maturation of the miRNA. This allows sets of nonoverlapping Morpholino oligos targeting a primary miRNA to be used as specificity controls; if two non-overlapping oligos targeting the same miRNA produce the same phenotype, this supports the hypothesis that the phenotype is due to knocking down the activity of the targeted miRNA and not due to an off-target effect. These techniques are explored in the following paper:

Kloosterman WP, Lagendijk AK, Ketting RF, Moulton JD, Plasterk RHA. Targeted inhibition of miRNA maturation with morpholinos reveals a role for miR-375 in pancreatic islet development. PLoS Biol. 2007;5(8): e203.

Available online at the PLoS Biology website.
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Old 01-24-2008, 05:06 PM
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Thumbs up Re: miRNA specificity controls: paper

Awesome paper, are you working on Morpholinos?

Also do you know if there are any new products/kits using this technology?
Is there increased efficiency other siRNA/RNAis?

very cool stuff
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Old 01-28-2008, 08:44 PM
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Default Re: miRNA specificity controls: paper

Hi Postdock,

Yes, I work for Gene Tools, so I work with Morpholinos. I'm a coauthor on the paper cited at the start of this thread.

For new products, if you are working with animals you might be interested in the Vivo-Morpholinos: www<dot>gene-tools<dot>com/vivomorpholinos

Regarding a comparison with siRNA, see this paper: www<dot>gene-tools<dot>com/files/Summerton2007siRNAcompare.pdf

Replace each <dot> with a "." to make the URLs work.
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