title: Activation of silenced cytokine gene promoters by the synergistic effect of TBP-TALE and VP64-TALE activators.
abstract: Recent work has shown that the combinatorial use of multiple TALE activators can selectively activate certain cellular genes in inaccessible chromatin regions. In this study, we aimed to interrogate the activation potential of TALEs upon transcriptionally silenced immune genes in the context of non-immune cells. We designed a unique strategy, in which a single TALE fused to the TATA-box binding protein (TBP-TALE) is coupled with multiple VP64-TALE activators. We found that our strategy is significantly more potent than multiple TALE activators alone in activating expression of IL-2 and GM-CSF in diverse cell origins in which both genes are otherwise completely silenced. Chromatin analysis revealed that the gene activation was due in part to displacement of a distinctly positioned nucleosome. These studies provide a novel epigenetic mechanism for artificial gene induction and have important implications for targeted cancer immunotherapy, DNA vaccine development, as well as rational design of TALE activators.
authors.affiliation: Department of Biology and Biochemistry, University of Houston, Houston, Texas, United States of America; Center for Nuclear Receptors and Cell Signaling, University of Houston, Houston, Texas, United States of America.
authors.forename: Kim
authors.initials: K
authors.lastname: Anthony
authors.affiliation: Department of Biology and Biochemistry, University of Houston, Houston, Texas, United States of America.
authors.forename: Abhijit
authors.initials: A
authors.lastname: More
authors.affiliation: Department of Biology and Biochemistry, University of Houston, Houston, Texas, United States of America; Center for Nuclear Receptors and Cell Signaling, University of Houston, Houston, Texas, United States of America.
authors.forename: Xiaoliu
authors.initials: X
authors.lastname: Zhang
mesh_terms: D001665:Binding Sites; D002460:Cell Line; D016207:Cytokines / Q000235:genetics*; D005786:Gene Expression Regulation; D023061:Gene Order; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics; D006801:Humans; D007376:Interleukin-2 / Q000235:genetics; D009928:Organ Specificity; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D035181:TATA-Box Binding Protein / Q000235:genetics / Q000378:metabolism*; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic; D015533:Transcriptional Activation
chemical_list: D016207:Cytokines; D007376:Interleukin-2; D011993:Recombinant Fusion Proteins; D035181:TATA-Box Binding Protein; D015534:Trans-Activators; D016178:Granulocyte-Macrophage Colony-Stimulating Factor
references.citation: J Leukoc Biol. 1995 May;57(5):767-73
references.pmid: 7759956
references.citation: Blood. 1995 Jan 1;85(1):80-6
references.pmid: 7528576
references.citation: Biotechnol Bioeng. 2013 Jul;110(7):1811-21
references.pmid: 23508559
references.citation: Nat Methods. 2013 Mar;10(3):239-42
references.pmid: 23377379
references.citation: Nucleic Acids Res. 2012 Jul;40(Web Server issue):W117-22
references.pmid: 22693217
references.citation: Cell Stem Cell. 2013 Feb 7;12(2):238-51
references.pmid: 23246482
references.citation: Curr Opin Microbiol. 2011 Feb;14(1):47-53
references.pmid: 21215685
references.citation: J Exp Med. 2003 Feb 17;197(4):413-23
references.pmid: 12591900
references.citation: Nat Biotechnol. 2013 Dec;31(12):1137-42
references.pmid: 24108092
references.citation: J Biol Chem. 1998 Jun 26;273(26):16509-16
references.pmid: 9632719
references.citation: J Biosci. 2003 Jun;28(4):423-36
references.pmid: 12799489
references.citation: Science. 2009 Dec 11;326(5959):1509-12
references.pmid: 19933107
references.citation: Int J Dev Biol. 2010;54(11-12):1589-96
references.pmid: 21404180
references.citation: Hum Gene Ther. 2012 Aug;23(8):883-90
references.pmid: 22587705
references.citation: J Immunol. 2011 Sep 15;187(6):3292-9
references.pmid: 21832163
references.citation: Mol Ther Nucleic Acids. 2013 Sep 03;2:e119
references.pmid: 24002729
references.citation: Mol Cell Biol. 2000 Jun;20(12):4350-8
references.pmid: 10825198
references.citation: J Immunol. 1999 Sep 15;163(6):3321-30
references.pmid: 10477602
references.citation: RNA. 2001 Nov;7(11):1578-88
references.pmid: 11720287
references.citation: Nucleic Acids Res. 2013 Oct;41(19):9197-207
references.pmid: 23921635
references.citation: Nucleic Acids Res. 2012 Nov;40(21):11163-72
references.pmid: 23019222
references.citation: J Immunol. 1999 Jul 1;163(1):120-9
references.pmid: 10384107
references.citation: Plant Mol Biol. 2012 Feb;78(3):311-21
references.pmid: 22167390
references.citation: Nucleic Acids Res. 2011 Jul;39(12):e82
references.pmid: 21493687
references.citation: Nat Methods. 2013 Mar;10(3):243-5
references.pmid: 23396285
references.citation: Nucleic Acids Res. 2012 Jul;40(12):5368-77
references.pmid: 22387464
references.citation: Nat Biotechnol. 2011 Feb;29(2):143-8
references.pmid: 21179091
references.citation: Genetics. 2010 Oct;186(2):757-61
references.pmid: 20660643
references.citation: Nat Biotechnol. 2013 Dec;31(12):1133-6
references.pmid: 24013198
references.citation: EMBO J. 1999 Sep 15;18(18):4969-80
references.pmid: 10487749
references.citation: Science. 2013 Feb 15;339(6121):816-9
references.pmid: 23413354
references.citation: Nature. 1994 Jun 9;369(6480):497-502
references.pmid: 8202141
references.citation: Science. 2009 Dec 11;326(5959):1501
references.pmid: 19933106
references.citation: J Cell Biochem. 2007 Apr 15;100(6):1477-92
references.pmid: 17133347
references.citation: Am J Physiol Lung Cell Mol Physiol. 2002 Aug;283(2):L460-7
references.pmid: 12114209
references.citation: Nat Commun. 2012 Jul 24;3:968
references.pmid: 22828628
references.citation: Proc Natl Acad Sci U S A. 1999 Mar 16;96(6):2668-73
references.pmid: 10077568
references.citation: J Immunol. 2002 Sep 1;169(5):2466-76
references.pmid: 12193716
references.citation: Nat Biotechnol. 2011 Feb;29(2):149-53
references.pmid: 21248753
references.citation: J Immunol. 2005 Apr 15;174(8):4639-46
references.pmid: 15814687
references.citation: PLoS One. 2011;6(5):e19509
references.pmid: 21625585
references.citation: PLoS One. 2012;7(7):e41734
references.pmid: 22860011
references.citation: J Interferon Cytokine Res. 2003 Feb;23(2):57-65
references.pmid: 12744771
references.citation: J Immunol. 2001 Oct 15;167(8):4494-503
references.pmid: 11591776
references.citation: Nature. 2012 Jan 18;483(7389):295-301
references.pmid: 22258509
references.citation: J Immunol. 2003 Apr 1;170(7):3724-31
references.pmid: 12646638
references.citation: Proc Natl Acad Sci U S A. 1997 Jul 8;94(14):7221-6
references.pmid: 9207072
references.citation: Mol Cell Biol. 2004 Sep;24(18):7914-30
references.pmid: 15340054
references.citation: Proc Natl Acad Sci U S A. 1996 Sep 3;93(18):9358-65
references.pmid: 8790334
references.citation: Biochem Biophys Res Commun. 2000 Mar 16;269(2):302-8
references.pmid: 10708547
references.citation: J Immunol. 2008 Jul 15;181(2):1272-81
references.pmid: 18606681
references.citation: Immunol Rev. 1998 Oct;165:287-300
references.pmid: 9850868
references.citation: Nucleic Acids Res. 2011 Nov;39(21):9283-93
references.pmid: 21813459
references.citation: Cell. 2013 Jan 31;152(3):642-54
references.pmid: 23333102