Knocking down miRNA using siRNA - (Aug/15/2006 )

Has anyone considered the possibility of knocking down an miRNA (at the pre-miRNA level) using siRNA? What are your thought on this idea?

-chicho-

People have used single stranded RNA to knockdown miRNA. A nature paper describes miRNA kockdown using "antagmirs".
http://www.ncbi.nlm.nih.gov/entrez/query.f...l=pubmed_docsum

There are also products available from Ambion to silence miRNA
http://www.ambion.com/catalog/ProdGrp.html...p;fkProdGrp=344

-pcrman-

Good references pcrman. As mentioned, this has successfully been done in the past. I've always thought of miRNA knowckdown as one of the more mind-bending experiment types...

-Elias-

QUOTE (pcrman @ Aug 15 2006, 06:29 PM)

Yes I know of such approaches. But, do to special circumstances I need to knockdown a microRNA at the pre-miRNA stage. Please don't ask the specifics of why I have to do it.
Thus, I ask if any of you think it would be feasible to use an siRNA to knockdown its precursor. I have doubt that such a thing could work due to the fact that the precursor is a stem-loop structure and the RISC complex might not be able to recognize it or even cut it.
Other collegues have suggested that it might be worth trying.

What are your opinions?

-chicho-

QUOTE (chicho @ Aug 16 2006, 01:56 PM)

I ask if any of you think it would be feasible to use an siRNA to knockdown its precursor. I have doubt that such a thing could work due to the fact that the precursor is a stem-loop structure and the RISC complex might not be able to recognize it or even cut it.
Other collegues have suggested that it might be worth trying.

I am very sure I read a paper published in Science or somewhere else using a synthetic siRNA targeting pre-miRNA loop and part of the stem structure to knockdown a miRNA but I couldn't find the reference after trying. I have actually had a siRNA targeting a miRNA synthesized but have not got a chance to test it.

Hope that helps.

-pcrman-

hi,
the paper you have been referring to was probably: J Biol Chem. 2005 Apr 29;280(17):16635-41. Epub 2005 Feb 18.
Depletion of human micro-RNA miR-125b reveals that it is critical for the proliferation of differentiated cells but not for the down-regulation of putative targets during differentiation.
Lee YS, Kim HK, Chung S, Kim KS, Dutta A.
they used si ran against the loop of the pre mir.
has anyone sucessfully worked with that so far?

best

-mitte03-