Controls in stable transfection experiments

ZhongLing Chai zhonglin.chai at med.monash.edu.au
Sun Feb 28 18:20:43 EST 1999


>I would suggest using the empty vector as a control. If you use the
>reverse orientation, isn't there the danger that the minus-strand RNA it
>makes would suppress translation of the endogenous gene, with who knows
>what effect?

Exactly. That's what we are trying to do: making an antisense construct (by
inserting the cDNA in an reverse orientation) allowing expressing of
anti-sense RNA, which, hopefully, will specifically (?) hybridize to the
endogenous mRNA and block the protein synthesis. By doing this, we want to
look at the effect on the cell cycle, of down-regulation of the protein we
are studying, although we don't know if this stragegy will work as well as
expected.




ZhongLin Chai, Ph.D.
____________________________________________
Department of Pathology and Immunology
Monash University Medical School
Alfred Hospital, Commercial Rd,
Prahran, Victoria, AUSTRALIA 3181
Telephone: (61 3) 9276 2698 (lab)
           (61 3) 9276 2696 (office)
Fax:       (61 3) 9529 6484
email:     zhonglin.chai at med.monash.edu.au
____________________________________________





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