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ARS Home » Midwest Area » Lexington, Kentucky » Forage-animal Production Research » Research » Publications at this Location » Publication #205129
Title: Applications of EcR Gene Switches Technology in Functional Genomics

Author
item TAVVA, VENKATA - UNIV OF KENTUCKY
item PALLI, SUBBA - UNIV OF KENTUCKY
item Dinkins, Randy
item COLLINS, GLENN - UNIV OF KENTUCKY

Submitted to: Archives of Insect Biochemistry and Physiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/10/2007
Publication Date: 6/24/2007
Citation: Tavva, V.S., Palli, S.R., Dinkins, R.D., Collins, G.B. 2007. Applications of EcR Gene Switches Technology in Functional Genomics. Archives of Insect Biochemistry and Physiology. 65:164-179. DOI: 10.1002/arch.20193.2007.

Interpretive Summary: Genetic engineering of plants through transgene technology is being used to enhance agronomic performance or improve quality traits in a wide variety of plant species, and has become a fundamental tool for basic research in plant biotechnology. Constitutive promoters are presently the primary means used to express transgenes in plants. Metabolic energy waste, negative pleiotropic effects and potential gene escape are some of the disadvantages associated with the use of constitutive promoters. Inducible gene regulation systems based on specific chemicals have many potential applications in agriculture and in the basic understanding of gene function. As a result several gene switches have been developed. However, the properties of the chemicals used in these switches make their use limited to research purposes. An ecdysone receptor gene switch is one of the best inducible gene regulation systems available, because the chemical, methoxyfenozide required for its regulation is registered for field use. An EcR gene switch with a potential for use in large-scale field applications and its applicability to a variety of plant species has been developed. The efficiency of EcR gene switches in inducing the target gene expression was also tested in functional genomic studies by regulating the expression of a Superman-like single zinc finger protein 11 (ZFP11) gene in both Arabidopsis and tobacco plants. The development of a highly sensitive and tightly regulated EcR gene switch along with other desirable properties such as availability of safe and field registered ligand should provide widespread use for this system.

Technical Abstract: Genetic engineering of plants through transgene technology is being used to enhance agronomic performance or improve quality traits in a wide variety of plant species, and has become a fundamental tool for basic research in plant biotechnology. Constitutive promoters are presently the primary means used to express transgenes in plants. However, inducible gene regulation systems based on specific chemicals have many potential applications in agriculture and in the basic understanding of gene function. As a result several gene switches have been developed. An ecdysone receptor gene switch is one of the best inducible gene regulation systems available, because the chemical, methoxyfenozide required for its regulation is registered for field use. An EcR gene switch with a potential for use in large-scale field applications and its applicability to a variety of plant species has been developed by adopting a two-hybrid format. In a two-hybrid switch format, the GAL4 DNA binding domain (GAL4 DBD) was fused to the ligand binding domain (LBD) of the Choristoneura fumiferana ecdysone receptor (CfEcR); and, the VP16 activation domain (VP16 AD) was fused to LBD of Locust migratoria retinoid x receptor (LmRXR). The sensitivity of the CfEcR gene switch was improved from micromolar to nanomolar concentrations of ligand by using the CfEcR:LmRXR two-hybrid switch. The efficiency of CfEcR:LmRXR two-hybrid gene switch in inducing the target gene expression was tested in functional genomic studies by regulating the expression of a Superman-like single zinc finger protein 11 (ZFP11) gene in both Arabidopsis and tobacco transgenic plants. The development of a sensitive EcR gene switch along with other desirable properties such as availability of safe and field registered ligand should provide widespread use for this system.