Can you transfect RNA into cells?

Jack M. Bernstein, M.D. BERNSTEIN at WSU-ID.DAYTON.OH.US
Sat Dec 9 23:49:21 EST 1995


Peter Wang <plw at mrc-lmb.cam.ac.uk> wrote:

>I was wondering about whether you can transfect RNA (produced in vitro 
>by T7) into eucaryotic cells by any of the methods used for DNA 
>(electroporation, lipofection, calcium phosphate, etc).
>If so, will the RNA be expressed (assuming it has appropriate 5'mG cap)?
>I know people have microinjected RNA into oocytes and had it expressed, 
>but I was thinking of garden-variety tissue-culture cells.
>Thanks for any insights!
>
>---------------------------------------------------------
>Peter Wang, M.D., Ph.D.
>MRC Centre for Protein Engineering,
>Hills Road, Cambridge, CB2 2QH, England
>
>Tel (01223) 402100  (international calls +44-1223-402100)
>    (01223) 402104  (direct number)
>Fax (01223) 402140
>---------------------------------------------------------
>
>
Absolutely. We are presently transfecting in the mRNA for T7
polymerase into eukaryotic cells and having enough T7 generated to
transcribe messages off a plasmid having a T7 promoter. Both the
plasmid with the T7 promoter AND the T7 messenger are transfected
TOGETHER in lipofectamine, a cationic liposome.

Please note that we have engineered an IRES site upstream of the T7pol
gene in order to have efficient translation of these uncapped
messages. References to this system can be found in many papers by
Bernie Moss.

I did some of this work with Andy Easton and Craig Pringle at the
University of Warwick while I was there on sabbatical. Please contact
myself, or Andy (AE at DNA.BIO.WARWICK.AC.UK) for further details. Good
luck.





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