siRNA - promoter compatibility

Chang Zhu czhu at changbioscience.com
Wed Jul 2 20:38:06 EST 2003


You might want to check 

http://www.changbioscience.com/stat/sirna.html

If you have designed your siRNA from scratch, there is a good chance
it wouldn't work. And the problem may be the choice of targets instead
of
promoter. 

Chang


"Peter Cherepanov" <peter.cherepanov at REMOVE.uz.kuleuven.ac.be> wrote in message news:<1056644473.858665 at seven.kulnet.kuleuven.ac.be>...
> that would be Northern.
> 
> You will easily find examples in literature.
> 
> I am just looking at a nice blot in:
> Nucleic Acids Res. 2003, 31(2):700-7.
> 
> you probably have seen this one:
> Science 2002, 296(5567):550-3.
> 
> Did you design your own hairpin siRNA from scratch? Have you tried a
> published one in the context of your vector?
> 
> Peter
> 
> 
> "Bertrand Collet" <b.collet at abdnnospam.ac.uk> wrote in message
> news:bdek43$5s6$1 at news.abdn.ac.uk...
> > Hi,
> >
> > I am actually making my own shRNA producing plasmid. I am working with
>  fish
> > cells and I made a construct with the fish U6 promoter.
> > When I use shRNA directed against luciferase (2 different shRNA made,
> > cotransfection with pCMV-FF luc using pRL as transfection control and
> > Dual-Glo assay kits), I don't get any significant silencing (so far).
> >
> > I would like to test wether my U6 promoter is active: is there a easy way
>  to
> > do that ? As it is a pol III promoter, no reporter genes could be used ...
> >
> > Sorry to highjack the thread !
> >
> > Thanks a lot in advance,
> >
> > Bertrand
> >
> > ""Peter Cherepanov"" <peter.cherepanov at uz.kuleuven.ac.be> wrote in message
> > news:003501c33b38$b3d69de0$9593a8c0 at uz.kuleuven.ac.be...
> > > 1) why not use human U6 promoter? It so easy easy to isolate it by PCR
> > >
> > > Check this:
> > >
> > > Qin XF, An DS, Chen IS, Baltimore D.Inhibiting HIV-1 infection in human
>  T
> > > cells by lentiviral-mediated delivery of small interfering RNA against
> > > CCR5.Proc Natl Acad Sci U S A. 2003 Jan 7;100(1):183-8.
> > >
> > > 2) mouse U6 promoter should work as well,
> > >
> > > Sui G, Soohoo C, Affar el B, Gay F, Shi Y, Forrester WC, Shi Y.A DNA
> > > vector-based RNAi technology to suppress gene expression in mammalian
> > > cells.Proc Natl Acad Sci U S A. 2002 Apr 16;99(8):5515-20.
> > >
> > > (there are plenty of examples using mouse U6 promoter in human cells,
>  check
> > > for uses of the pmU6 vector)
> > >
> > >
> > >
> > > Peter
> > >
> > >
> > >
> > >
> > >
> > >
> > >
> > > ---
> >
> >
> >



More information about the Methods mailing list