Transcription vectors for RNAi?

Colin Dolphin colin.dolphin at kcl.ac.uk
Mon Jun 19 08:42:37 EST 2000


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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