Increment of mRNA level after siRNA

Yvonne Couch via methods%40net.bio.net (by yvonne.couch from dpag.ox.ac.uk)
Thu Jan 31 11:54:35 EST 2008


There have been a couple of papers that suggest that targeting specific
regions of the gene, usually the 3'UTR can actually cause upregulation of
genes rather than down regulation.  This may be a possible explanation of
what you have inadvertently done.  I can't find the paper that did this
using siRNA/plasmids but if you read this months Nature Methods there's a
paper on micro RNA's upregulating genes dependent on cell status.
Hope this help?
Y.

-----Original Message-----
From: methods-bounces from oat.bio.indiana.edu
[mailto:methods-bounces from oat.bio.indiana.edu] On Behalf Of Kamalian, Laleh
Sent: 31 January 2008 14:25
To: methods
Subject: Increment of mRNA level after siRNA

 
Hi
Thanks to people who helped me last time.  This time its not a desperate
question.  Only to ask your opinions.  I have successfully used a plasmid
vector to subside the mRNA level of the gene I'm working on.  I used 4
different oligonucleotides to insert into the plasmid and used them to
target my gene.  Although most of the plasmids have successfully decreased
the mRNA level (different levels of course), one of the clones has actually
increased the level of the gene's mRNA.  I can't explain it.  Some of my
colleagues suggest the compensatory mechanisms of the cell.  I think its
simply not being knocked down or maybe an error in real time PCR? I had seen
this before working on another cell line.  What do you think?
LK
 
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