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Journal of Virology, May 2000, p. 4679-4687, Vol. 74, No. 10
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
The cHS4 Insulator Increases the Probability of
Retroviral Expression at Random Chromosomal Integration Sites
Stefano
Rivella,
John A.
Callegari,
Chad
May,
Cui Wen
Tan, and
Michel
Sadelain*
Department of Human Genetics, Memorial
Sloan-Kettering Cancer Center, New York, New York 10021
Received 5 October 1999/Accepted 4 February 2000
Retroviruses are highly susceptible to transcriptional silencing
and position effects imparted by chromosomal sequences at their
integration site. These phenomena hamper the use of recombinant retroviruses as stable gene delivery vectors. As insulators are able to
block promoter-enhancer interactions and reduce position effects in
some transgenic animals, we examined the effect of an insulator on the
expression and structure of randomly integrated recombinant
retroviruses. We used the cHS4 element, an insulator from the chicken
-like globin gene cluster, which has been shown to reduce position
effects in transgenic Drosophila. A large panel of mouse
erythroleukemia cells that bear a single copy of integrated recombinant
retroviruses was generated without using drug selection. We show that
the cHS4 increases the probability that integrated proviruses will
express and dramatically decreases the level of de novo methylation of
the 5' long terminal repeat. These findings support a primary role of
methylation in the silencing of retroviruses and suggest that cHS4
could be useful in gene therapy applications to overcome silencing of
retroviral vectors.
*
Corresponding author. Mailing address: Department of
Human Genetics, Box 182, Memorial Sloan-Kettering Cancer Center, 1275 York Ave., New York, NY 10021. Phone: (212) 639-6190. Fax: (917) 432-2340. E-mail: m-sadelain{at}ski.mskcc.org.
Journal of Virology, May 2000, p. 4679-4687, Vol. 74, No. 10
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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