siliencing mulitiple genes by RNAi

Ricky Boernke ricky_boernke at gmx.net
Fri Jun 27 04:47:41 EST 2003


We have fused together two sequences and both genes were targeted
with similar efficiency. More should also be possible.

Ricky


s.xiao at uea.ac.uk ("Shunyuan Xiao") wrote in message 
news:<001801c32f71$428fb4f0$0b41de8b at TURNERXP2>...
>  What about making a chimeric RNAi construct:  linking  unique sequences from
>  each of  3 or 4 or more target genes first and then making the inverted
>  repeat?   Theroetically this kind of construct should generate 100% matching
>  & unique small double stranded RNA effective in silencing the corresponding
>  genes.
>
>  Has anybody done this and got it work??
>
>
>  Shunyuan Xiao, PhD
>
>  School of Biological Sciences
>
>  University of East Anglia, Norwich  NR4 7TJ
>
>  United Kingdom
>
>
>
>
>
>  ----- Original Message -----
>  From: ""Michael L. Sullivan"" <mlsulliv at facstaff.wisc.edu>
>  Newsgroups: bionet.genome.arabidopsis
>  To: <arab-gen at net.bio.net>
>  Sent: Monday, June 09, 2003 5:47 PM
>  Subject: Re: siliencing mulitiple genes by RNAi
>
>
>  >  <x-flowed> From my reading of the literature, I think it should be
>   possible
>  >  provided there is sufficient sequence identity among the genes.  The
>  >  figure 85% sticks out in my mind for plants, although I don't recall
>  >  which paper I got that from (perhaps from something from Peter
>  >  Waterhouse's lab).
>  >
>  >  Depending on how much similarity there is between the genes, you
>  >  might feel more or less confident about taking a "one construct"
>  >  approach.  If the similarity among genes were >90%, I'd be tempted to
>  >  just make one construct.  Less than that I might hedge my bets with
>  >  multiple constructs.  But first I'd scour the literature a bit more--
>  >  I think the data is out there.
>  >
>  >  Good luck.
>  >
>  >  Mike
>  >
>  >
>  >
>  >  >Has anyone any experience of silencing multiple genes with the same
>  >  >construct? Can it be done for 3 genes, or will I have to make three
>  >  >separate knockouts and cross them?
>  >  >
>  >  >Lee
>  >  >
>  >  >---
>  >
>  >
>  >  --
>  >
>  >  Michael L. Sullivan, Ph.D
>  >  Research Plant Molecular
>  >  Geneticist
>  >  U.S. Dairy Forage Research Center
>  >  1925 Linden Drive West
>  >  Madison WI, 53706
>  >
>  >  (608) 264-5397 Phone
>  >  (608) 264-5147 Fax
>  >  ---
>  >
>  >  </x-flowed>
>  >
>
>  ---
---



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