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Reynolds, Lindsey, Ullman, Christopher, Moore, Michael, Isalan, Mark, West, Michelle J, Clapham, Paul, Klug, Aaron and Choo, Yen (2003) Repression of the HIV-1 5' LTR promoter and inhibition of HIV-1 replication by using engineered zinc-finger transcription factors. Proceedings of the National Academy of Sciences, 100 (4). pp. 1615-1620. ISSN 0027-8424
Full text not available from this repository.Abstract
Zinc finger domains are small DNA-binding modules that can be engineered to bind desired target sequences. Functional transcription factors can be made from these DNA-binding modules, by fusion with an appropriate effector domain. In this study, eight three-zinc-finger proteins (ZFPs) that bound HIV-1 sequences in vitro were engineered into transcription repressors by linking them to the Krüppel-associated box (KRAB) repressor domain (KOX1). One protein, ZFP HIVB-KOX, which bound to a 9-bp region overlapping two Sp1 sites, was found to repress a Tat-activated 5' LTR cellular HIV-reporter assay to almost basal levels. A related six-finger protein, HIVBA'-KOX, was made to target all three Sp1 sites in the 5' LTR promoter and efficiently inhibited both basal and Tat-activated transcription in unstimulated and mitogen-stimulated T cells. In contrast, a combination of two unlinked three-finger ZFPs, HIVA'-KOX and HIVB-KOX, which bind over the same region of DNA, resulted in less effective repression. Finally, HIVBA'-KOX was tested for its capacity to block viral replication in a cellular infection assay using the HIV-1 HXB2 strain. This ZFP was found to inhibit HIV-1 replication by 75% compared with control constructs, thus demonstrating the potential of this approach for antiviral therapy.
| Item Type: | Article |
|---|---|
| Additional Information: | West generated 30% of the data in this paper as a collaboration and co-wrote the paper. Reynolds and Ullman are joint first author so West is fourth author. West's data formed a key part of this paper describing novel effective HIV-1 inhibitors.Reynolds and Ullman are joint first authors. West is fourth but not appearing on the list of authors at all so looks like no internal authors. |
| Schools and Departments: | School of Life Sciences > Biochemistry |
| Depositing User: | Michelle West |
| Date Deposited: | 06 Feb 2012 19:50 |
| Last Modified: | 04 May 2012 08:48 |
| URI: | http://sro.sussex.ac.uk/id/eprint/22407 |