Drug-inducible and Simultaneous Regulation of Endogenous Genes by Single-chain Nuclear Receptor-based Zinc-finger Transcription Factor Gene Switches

Overview

Journal Gene Ther

Date 2008 Jun 6

PMID 18528430

Citations 14

Authors

L Magnenat

L J Schwimmer

C F Barbas 3rd

Affiliations

Soon will be listed here.

Abstract

Chemically inducible gene switches that regulate expression of endogenous genes have multiple applications for basic gene expression research and gene therapy. Single-chain zinc-finger transcription factors that utilize either estrogen receptor homodimers or retinoid X receptor-alpha/ecdysone receptor heterodimers are shown here to be effective regulators of ICAM-1 and ErbB-2 transcription. Using activator (VP64) and repressor (Krüppel-associated box) domains to impart regulatory directionality, ICAM-1 was activated by 4.8-fold and repressed by 81% with the estrogen receptor-inducible transcription factors. ErbB-2 was activated by up to threefold and repressed by 84% with the retinoid X receptor-alpha/ecdysone receptor-inducible transcription factors. The dynamic range of these proteins was similar to the constitutive system and showed negligible basal regulation when ligand was not present. We have also demonstrated that the regulation imposed by these inducible transcription factors is dose dependent, sustainable for at least 11 days and reversible upon cessation of drug treatment. Importantly, these proteins can be used in conjunction with each other with no detectable overlap of activity enabling concurrent and temporal regulation of multiple genes within the same cell. Thus, these chemically inducible transcription factors are valuable tools for spatiotemporal control of gene expression that should prove valuable for research and gene therapy applications.

Citing Articles

Genome Editing and Diabetic Cardiomyopathy.

Kambis T, Mishra P Adv Exp Med Biol. 2022; 1396:103-114.

PMID: 36454462 PMC: 10155862. DOI: 10.1007/978-981-19-5642-3_7.

Integrated analysis of transcription factors and targets co-expression profiles reveals reduced correlation between transcription factors and target genes in cancer.

Liang J, Cui Y, Meng Y, Li X, Wang X, Liu W Funct Integr Genomics. 2018; 19(1):191-204.

PMID: 30251028 DOI: 10.1007/s10142-018-0636-6.

Genome-Editing Technologies: Principles and Applications.

Gaj T, Sirk S, Shui S, Liu J Cold Spring Harb Perspect Biol. 2016; 8(12).

PMID: 27908936 PMC: 5131771. DOI: 10.1101/cshperspect.a023754.

Targeted epigenetic repression of a lymphoma oncogene by sequence-specific histone modifiers induces apoptosis in DLBCL.

Luo H, Schmidt J, Lee Y, Oltz E, Payton J Leuk Lymphoma. 2016; 58(2):445-456.

PMID: 27268204 PMC: 5206989. DOI: 10.1080/10428194.2016.1190973.

Editing the epigenome: technologies for programmable transcription and epigenetic modulation.

Thakore P, Black J, Hilton I, Gersbach C Nat Methods. 2016; 13(2):127-37.

PMID: 26820547 PMC: 4922638. DOI: 10.1038/nmeth.3733.

References

Xu D, Ye D, Fisher M, Juliano R . Selective inhibition of P-glycoprotein expression in multidrug-resistant tumor cells by a designed transcriptional regulator. J Pharmacol Exp Ther. 2002; 302(3):963-71. DOI: 10.1124/jpet.102.033639. View

Blancafort P, Magnenat L, Barbas 3rd C . Scanning the human genome with combinatorial transcription factor libraries. Nat Biotechnol. 2003; 21(3):269-74. DOI: 10.1038/nbt794. View

Dreier B, Beerli R, Segal D, Flippin J, Barbas 3rd C . Development of zinc finger domains for recognition of the 5'-ANN-3' family of DNA sequences and their use in the construction of artificial transcription factors. J Biol Chem. 2001; 276(31):29466-78. DOI: 10.1074/jbc.M102604200. View

Park K, Lee D, Lee H, Lee Y, Jang Y, Kim Y . Phenotypic alteration of eukaryotic cells using randomized libraries of artificial transcription factors. Nat Biotechnol. 2003; 21(10):1208-14. DOI: 10.1038/nbt868. View

Harwerth I, Wels W, Marte B, Hynes N . Monoclonal antibodies against the extracellular domain of the erbB-2 receptor function as partial ligand agonists. J Biol Chem. 1992; 267(21):15160-7. View

Bartsevich V, Juliano R . Regulation of the MDR1 gene by transcriptional repressors selected using peptide combinatorial libraries. Mol Pharmacol. 2000; 58(1):1-10. DOI: 10.1124/mol.58.1.1. View

Beerli R, Schopfer U, Dreier B, Barbas 3rd C . Chemically regulated zinc finger transcription factors. J Biol Chem. 2000; 275(42):32617-27. DOI: 10.1074/jbc.M005108200. View

Margolin J, Friedman J, Meyer W, Vissing H, Thiesen H, Rauscher 3rd F . Krüppel-associated boxes are potent transcriptional repression domains. Proc Natl Acad Sci U S A. 1994; 91(10):4509-13. PMC: 43815. DOI: 10.1073/pnas.91.10.4509. View

Rebar E, Huang Y, Hickey R, Nath A, Meoli D, Nath S . Induction of angiogenesis in a mouse model using engineered transcription factors. Nat Med. 2002; 8(12):1427-32. DOI: 10.1038/nm1202-795. View

10.

Liu Q, Segal D, Ghiara J, Barbas 3rd C . Design of polydactyl zinc-finger proteins for unique addressing within complex genomes. Proc Natl Acad Sci U S A. 1997; 94(11):5525-30. PMC: 20811. DOI: 10.1073/pnas.94.11.5525. View

11.

Toniatti C, Bujard H, Cortese R, Ciliberto G . Gene therapy progress and prospects: transcription regulatory systems. Gene Ther. 2004; 11(8):649-57. DOI: 10.1038/sj.gt.3302251. View

12.

Beerli R, Barbas 3rd C . Engineering polydactyl zinc-finger transcription factors. Nat Biotechnol. 2002; 20(2):135-41. DOI: 10.1038/nbt0202-135. View

13.

Beerli R, Dreier B, Barbas 3rd C . Positive and negative regulation of endogenous genes by designed transcription factors. Proc Natl Acad Sci U S A. 2000; 97(4):1495-500. PMC: 26462. DOI: 10.1073/pnas.040552697. View

14.

Segal D, Dreier B, Beerli R, Barbas 3rd C . Toward controlling gene expression at will: selection and design of zinc finger domains recognizing each of the 5'-GNN-3' DNA target sequences. Proc Natl Acad Sci U S A. 1999; 96(6):2758-63. PMC: 15842. DOI: 10.1073/pnas.96.6.2758. View

15.

Jamieson A, Miller J, Pabo C . Drug discovery with engineered zinc-finger proteins. Nat Rev Drug Discov. 2003; 2(5):361-8. DOI: 10.1038/nrd1087. View

16.

Zhang L, Spratt S, Liu Q, Johnstone B, Qi H, Raschke E . Synthetic zinc finger transcription factor action at an endogenous chromosomal site. Activation of the human erythropoietin gene. J Biol Chem. 2000; 275(43):33850-60. DOI: 10.1074/jbc.M005341200. View

17.

Dreier B, Segal D, Barbas 3rd C . Insights into the molecular recognition of the 5'-GNN-3' family of DNA sequences by zinc finger domains. J Mol Biol. 2000; 303(4):489-502. DOI: 10.1006/jmbi.2000.4133. View

18.

Kang J, Kim J . Zinc finger proteins as designer transcription factors. J Biol Chem. 2000; 275(12):8742-8. DOI: 10.1074/jbc.275.12.8742. View

19.

Ren D, Collingwood T, Rebar E, Wolffe A, Camp H . PPARgamma knockdown by engineered transcription factors: exogenous PPARgamma2 but not PPARgamma1 reactivates adipogenesis. Genes Dev. 2002; 16(1):27-32. PMC: 155307. DOI: 10.1101/gad.953802. View

20.

Ayer D, Laherty C, Lawrence Q, Armstrong A, Eisenman R . Mad proteins contain a dominant transcription repression domain. Mol Cell Biol. 1996; 16(10):5772-81. PMC: 231578. DOI: 10.1128/MCB.16.10.5772. View