antisense technology?

Elizabeth Rosenberg erosenbe at gpu2.srv.ualberta.ca
Thu Nov 21 17:09:27 EST 1996


Hi!

Our group is about to embark on the glorious journey of downregulation of
a gene by use of antisense constructs.  We are very new to this, and would
appreciate any and all feedback anyone might have.  Especially, we are
interested in knowing what the sources for "standard" protocols for this
technology are these days.  (Other than Maniatis or the red book).

Specifically, we have inserted our gene of interest into the pREP9 vector,
transfected this construct into a CHO mutant cell line that is defective
in the gene of interest, and showed restoration of the "normal" phenotype.
Yea, yippee, hurrah, so far, so good.  

Now we would like to insert an antisense fragment of the same gene into
the pREP10 vector and tranfect this construct into "normal" CHO cells that
are the "parent" of our mutant strain.

Does this strategy seem reasonable?  What else do we need to consider?
What are the caveats, etc. from those in the know?

I have already searched the archives of this newsgroup, and information on
this topic is scanty to say the least.  Again, any advice, and especially
suggestions of good sources of information would be most appreciated.

Thanks in advance,

Beth
_____________________
Beth Rosenberg                    erosenbe at gpu.srv.ualberta.ca
Cross Cancer Institute            
University of Alberta		 "The sheep and the wolf do not
                                 agree on the definition of liberty" 
				            -Carl Sandburg



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