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gene therapy question

Olav Hungnes ohungnes at bioslave.uio.no
Thu May 26 03:19:48 EST 1994

Dr. J.P. Clewley (jclewley at crc.ac.uk) wrote:
: In article <2rrmho$pcd at news.u.washington.edu>, frv at u.washington.edu (Franklin Vincenzi) writes:
: > Dr. J.P. Clewley <jclewley at crc.ac.uk> wrote:
: > 
: > Some helpful advice, and then
: > 
: > >What does anyone else think?
: > 
: > I think I'm glad I asked what the lot of you think about this!  Thanks 
: > for the crash course.
: > 
: > Regards, Franklin

: One thing I forgot to mention is that its more difficult to engineer
: a negative strand virus than a positive strand one. If its +ve, you
: can make a cDNA clone & transfect it in cells & recover progeny virus.
: Works for alphaviruses (eg. Sindbis) picornaviruses (e.g. polio),
: retroviruses (e.g. HIV). Recovering -ve strand virus from transfected
: DNA has not been done reproducibly, as far as I know. Is there a
: confirmed example I'm missing?

: Cheers, Jon

Peter Palese's group has developed a method to rescue gene-manipulated 
influenza virus segments (negative-strand, multi-segmented). They make 
runoff transcripts in the presence of virus nucleoprotein to obtain minus 
strand RNA that can be transfected into infected cells. You then have to 
find a way to select/screen for the desired reassortant virus. Certainly 
more work than with +strand virus.

Olav Hungnes                     ohungnes at embnet.uio.no
National Institute               Phone  (+47)22042200
of Public Health                 FAX    (+47)22353605

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