un691cs at genius.embnet.dkfz-heidelberg.de
Thu Nov 28 04:42:21 EST 1996
In article <57e912$jfb at kadri.ut.ee>, mspeek at tamm.eenet.ee says...
>Elizabeth Rosenberg (erosenbe at gpu2.srv.ualberta.ca) wrote:
>: 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.
>: 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.
>I would not recommend antisense experiments such as yours. This is because
>it is remarkably inefficient (for review see NAR 21:4383-91 (1993))
>I spent 1.5 yrs to study RNA-RNA hybridization/annealing between
>a pair of naturally occurring sense-antisense mRNAs, but found
>very little of them in duplex (Mol. Endo.9:1655-65 (1995))
>Mart (mspeek at ebc.ee in Estonia)
I would also think that you may need several, if not many very good controls.
I have seen reports that sense oligonucleotides for instance may have stronger
inhibitory effects than the antisense ! so one control will clearly not do.
Did you ever think about using a hammerhead ribozyme structure a la Cech,
or does anybody out there have experience with it ? Last time I heard about
hammerheads, the results were also quit disputable though... seems that it
doesn't work always neither...
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