title: A combinatorial bidirectional and bicistronic approach for coordinated multi-gene expression in corn.
abstract: Transgene stacking in trait development process through genetic engineering is becoming complex with increased number of desired traits and multiple modes of action for each trait. We demonstrate here a novel gene stacking strategy by combining bidirectional promoter (BDP) and bicistronic approaches to drive coordinated expression of multi-genes in corn. A unidirectional promoter, Ubiquitin-1 (ZMUbi1), from Zea mays was first converted into a synthetic BDP, such that a single promoter can direct the expression of two genes from each end of the promoter. The BDP system was then combined with a bicistronic organization of genes at both ends of the promoter by using a Thosea asigna virus 2A auto-cleaving domain. With this gene stacking configuration, we have successfully obtained expression in transgenic corn of four transgenes; three transgenes conferring insect (cry34Ab1 and cry35Ab1) and herbicide (aad1) resistance, and a phiyfp reporter gene using a single ZMUbi1 bidirectional promoter. Gene expression analyses of transgenic corn plants confirmed better coordinated expression of the four genes compared to constructs driving each gene by independent unidirectional ZmUbi1 promoter. To our knowledge, this is the first report that demonstrates application of a single promoter for co-regulation of multiple genes in a crop plant. This stacking technology would be useful for engineering metabolic pathways both for basic and applied research.
authors: Dow AgroSciences LLC, 9330 Zionsville Road, Indianapolis, IN, 46268, USA, Skumar9@dow.com.
authors: Sandeep
authors: S
authors: Kumar
authors: Diaa
authors: D
authors: AlAbed
authors: John T
authors: JT
authors: Whitteck
authors: Wei
authors: W
authors: Chen
authors: Sara
authors: S
authors: Bennett
authors: Andrew
authors: A
authors: Asberry
authors: Xiujuan
authors: X
authors: Wang
authors: Daniel
authors: D
authors: DeSloover
authors: Murugesan
authors: M
authors: Rangasamy
authors: Terry R
authors: TR
authors: Wright
authors: Manju
authors: M
authors: Gupta
mesh_terms: D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D003313:Zea mays / Q000235:genetics*
references: Plant Physiol. 2004 May;135(1):16-24
references: 15141063
references: Plant Cell. 1989 Jan;1(1):115-22
references: 2535459
references: Genes Dev. 2002 Oct 15;16(20):2583-92
references: 12381658
references: Plant Physiol. 2003 Jun;132(2):988-98
references: 12805627
references: Plant Biotechnol J. 2010 Oct;8(8):928-38
references: 20649940
references: J Exp Bot. 2005 Sep;56(419):2345-53
references: 16014367
references: J Integr Plant Biol. 2012 Apr;54(4):250-9
references: 22348899
references: J Econ Entomol. 2012 Oct;105(5):1825-34
references: 23156183
references: Proc Natl Acad Sci U S A. 2010 Nov 23;107(47):20240-5
references: 21059954
references: GM Crops. 2010 Jul-Sep;1(4):220-9
references: 21844677
references: Proc Natl Acad Sci U S A. 1989 May;86(9):3219-23
references: 16594033
references: Trends Plant Sci. 2007 Mar;12(3):118-24
references: 17292658
references: J Biotechnol. 2006 May 3;123(1):1-12
references: 16324763
references: Biomed Res Int. 2013;2013:291730
references: 23878801
references: EMBO J. 1987 Dec 20;6(13):3901-7
references: 3327686
references: Plant Biotechnol J. 2005 Mar;3(2):141-55
references: 17173615
references: PLoS One. 2013;8(1):e53079
references: 23308139
references: Biotechnol J. 2010 Feb;5(2):213-23
references: 19946875
references: Curr Opin Biotechnol. 2002 Apr;13(2):136-41
references: 11950565
references: Mol Ther. 2005 Sep;12(3):569-74
references: 15964244
references: Nat Biotechnol. 2001 Jul;19(7):677-9
references: 11433282
references: Plant Cell. 2007 Mar;19(3):943-58
references: 17384170
references: J Gen Virol. 2001 May;82(Pt 5):1013-25
references: 11297676
references: Mol Biol Evol. 2004 May;21(5):841-50
references: 14963095
references: Nat Biotechnol. 1996 Oct;14(10):1274-8
references: 9631092
references: Biotechnol Lett. 2011 Feb;33(2):387-93
references: 20953667
references: Planta. 2007 Jan;225(2):365-79
references: 16924537
references: BMC Biotechnol. 2013 Aug 22;13:67
references: 23968294
references: EMBO J. 1984 Dec 1;3(12):2723-30
references: 16453574
references: J Econ Entomol. 2005 Feb;98(1):1-8
references: 15765660
references: J Gen Virol. 1999 Jul;80 ( Pt 7):1855-63
references: 10423156
references: Plant Mol Biol. 2010 Jul;73(4-5):363-78
references: 20354894
references: Planta. 2001 Sep;213(5):731-40
references: 11678277