Replies to post #9509 on Cytogenix (fka CYGX)

[Tatonkano62]

Found this; it includes Peter Glazer

http://www.nature.com/cgi-taf/DynaPage.taf?file=/ng/journal/v20/n2/abs/ng1098_212.html&dynoption...

Targeted gene knockout mediated by triple helix forming oligonucleotides

Alokes Majumdar1, Alexander Khorlin2, Natalia Dyatkina2, F.-L. Michael Lin2, James Powell4, Jilan Liu2, Zhizhong Fei2, Yuri Khripine2, Kyoichi A. Watanabe2, Jay George2, Peter M. Glazer3 & Michael M. Seidman1


Triple helix forming oligonucleotides (TFOs) recognize and bind sequences in duplex DNA and have received considerable attention because of their potential for targeting specific genomic sites1-3. TFOs can deliver DNA reactive reagents to specific sequences in purified chromosomal DNA (ref. 4) and nuclei5. However, chromosome targeting in viable cells has not been demonstrated, and in vitro experiments indicate that chromatin structure is incompatible with triplex formation6-8. We have prepared modified TFOs, linked to the DNA-crosslinking reagent psoralen, directed at a site in the Hprt gene. We show that stable Hprt-deficient clones can be recovered following introduction of the TFOs into viable cells and photoactivation of the psoralen. Analysis of 282 clones indicated that 85% contained mutations in the triplex target region. We observed mainly deletions and some insertions. These data indicate that appropriately constructed TFOs can find chromosomal targets, and suggest that the chromatin structure in the target region is more dynamic than predicted by the in vitro experiments.



1. National Institute on Aging, NIH, 5600 Nathan Shock Dr., Baltimore, Maryland 21224-6825, USA.
2. Codon Pharmaceuticals, 200 Perry Parkway, Gaithersburg, Maryland 20877, USA.
3. Department of Therapeutic Radiology, Yale University, New Haven, Connecticut 06510, USA.
4. Ribozyme Pharmaceuticals Inc., Boulder, Colorado, USA.
Correspondence should be addressed to M M Seidman. e-mail: [e-mail]

[schrodbox]

You stole my thunder!