Transcription vectors for RNAi?

Nobody nobody at hgmp.mrc.ac.uk
Sat Jun 24 12:41:29 EST 2000


 Hi Colin,

    The first part of your question, and the approach you propose, are good
thinking but someone did think about it before you... Here's the reference:

Nature Genetics volume 24, page 180-183, february issue 2000.

=46or the last part, I think you may get some light by reading the original
paper by Fire et al.:

=46ire A, Xu S, Montgomery MK, Kostas SA, Driver SE, Mello CC.
Potent and specific genetic interference by double-stranded RNA in
Caenorhabditis elegans.
Nature. 1998 Feb 19;391(6669):806-11.

Good luck.

Tom

_____________

Thomas Duchaine
PhD Student, Molecular Biology Program
Biochemistry Dep.
University of Montreal
_____________

Colin Dolphin <colin.dolphin at kcl.ac.uk> a =C8crit dans le message :
8ilnm0$r9m$1 at mercury.hgmp.mrc.ac.uk...
> We're planning to construct vectors for transcription of sense and
> antisense RNAs for RNAi. To simplify the transcription process would it
> be possible to construct a single vector containing two copies of the
> insert in appropriate orientations such that transcription from one end
> would produce a single transcript that would then fold to give a dsRNA
> molecule with a short loop at one end? This would seem to be a more
> straightfoward method of generating dsRNA than doing two separate
> transcription reactions, quantifying yield and then mixing sense and
> antisense together. Also, how long should the dsRNA molecule be to effect
> silencing? Could we, for instance, PCR an exon from genomic as the
> template for transcription?
>
> Colin
>
> Colin Dolphin
> Dept of Pharmacy
> Kings College London
> London SE1 8WA
> colin.dolphin at kcl.ac.uk
>
>


---


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