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Biotech / Medical : Sangamo Therapeutics, Inc. SGMO

SGMO 0.530

+7.5%

Dec 4 3:59 PM EST

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To: Mike McFarland who started this subject	7/27/2000 4:47:01 PM
From: sim1	Read Replies (1) of 368

From Yahoo. messages.yahoo.com Synthetic Zinc Finger Transcription Factor Action at an Endogenous Chromosomal Site: Activation of the Human Erythropoietin Gene Lei Zhang, S. Kaye Spratt, Qiang Liu, Brian Johnstone, Hong Qi, Eva E. Raschke, Andrew C.Jamieson, Edward J.Rebar, Alan P.Wolffe* and Casey C.Case Sangamo BioSciences Inc. Point Richmond Tech Center 501 Canal Blvd., Suite A100 Richmond CA 94804 *Corresponding author Phone: (510) 970-6000 Ext 216 Fax: (510) 236-8951 awolffe@sangamo.com Copyright 2000 by The American Society for biochemistry and Molecular Biology, Inc. Running title : Transcriptional activation of chromatin ABSTRACT We have targeted the activation of an endogenous chromosomal locus including the human erythropoietin gene using synthetic transcription factors. These transcription factors are targeted to particular DNA sequences in the 5 flanking region of the erythropoietin gene through engineering of a zinc finger DNA binding domain. The DNA binding domain is linked to a VP16 transcriptional activation domain. We find that these synthetic transcription factors invariably activate transiently transfected templates in which sequences within the 5 flank of the erythropoietin gene are fused to a luciferase reporter. The efficiency of activation under these circumstances at a defined site is dependent on DNA binding affinity. In contrast, only a subset of these same zinc finger proteins is able to activate the endogenous chromosomal locus. The activity of these proteins is influenced by their capacity to gain access to their recognition elements within the chromatin infrastructure. Zinc finger transcription factors will provide a powerful tool to probe the determinants of chromatin accessibility and remodeling within endogenous chromosomal loci. jbc.org

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