Antisense Design

[Jack M. Bernstein, M.D.]

This is a general posting.

We have been playing with anti-sense oligos for about 18 months. Our goal is 
to inhibit Respiratory Syncytial Virus, a negative stranded RNA virus which is 
completely cytoplasmic in its replicative cycle.

We have discovered that either native DNA oligos or phosphorothioate 
derivatives seem to do much better vis a vis cell penetration when they are 
mixed with lipofectamine. Unfortunately, they probably transit rapidly to the 
nucleus where they are unavailable to inhibit our virus.

Three general questions:

Does anyone have a feeling if conjugation of an oligo to cholesterol or 
poly-L-lysine will help penetration? The literature supports this notion.

If we assume that they will poenetrate better as a cholesterol or 
poly-L-lysine conjugate, which would be preferable?

Finally, will either poly-L-lysine or cholesterol help the oligo to remain in 
the cytoplasm for a prolonged period of time or will the oligo naturally 
compartmentalize to the nucleus?

Thanks for anyones input. Perhaps this will start an "anti-sense thread". I 
haven't seen much on it in this newsgroup.

Jack Bernstein