title: Transcriptional activation of Brassica napus β-ketoacyl-ACP synthase II with an engineered zinc finger protein transcription factor.
abstract: Targeted gene regulation via designed transcription factors has great potential for precise phenotypic modification and acceleration of novel crop trait development. Canola seed oil composition is dictated largely by the expression of genes encoding enzymes in the fatty acid biosynthetic pathway. In the present study, zinc finger proteins (ZFPs) were designed to bind DNA sequences common to two canola β-ketoacyl-ACP Synthase II (KASII) genes downstream of their transcription start site. Transcriptional activators (ZFP-TFs) were constructed by fusing these ZFP DNA-binding domains to the VP16 transcriptional activation domain. Following transformation using Agrobacterium, transgenic events expressing ZFP-TFs were generated and shown to have elevated KASII transcript levels in the leaves of transgenic T(0)  plants when compared to 'selectable marker only' controls as well as of T(1)  progeny plants when compared to null segregants. In addition, leaves of ZFP-TF-expressing T(1)  plants contained statistically significant decreases in palmitic acid (consistent with increased KASII activity) and increased total C18. Similarly, T(2)  seed displayed statistically significant decreases in palmitic acid, increased total C18 and reduced total saturated fatty acid contents. These results demonstrate that designed ZFP-TFs can be used to regulate the expression of endogenous genes to elicit specific phenotypic modifications of agronomically relevant traits in a crop species.
authors.affiliation: Dow AgroSciences, Indianapolis, IN, USA.
authors.forename: Manju
authors.initials: M
authors.lastname: Gupta
authors.forename: Russell C
authors.initials: RC
authors.lastname: DeKelver
authors.forename: Asha
authors.initials: A
authors.lastname: Palta
authors.forename: Carla
authors.initials: C
authors.lastname: Clifford
authors.forename: Sunita
authors.initials: S
authors.lastname: Gopalan
authors.forename: Jeffrey C
authors.initials: JC
authors.lastname: Miller
authors.forename: Stephen
authors.initials: S
authors.lastname: Novak
authors.forename: Daniel
authors.initials: D
authors.lastname: Desloover
authors.forename: Daniel
authors.initials: D
authors.lastname: Gachotte
authors.forename: James
authors.initials: J
authors.lastname: Connell
authors.forename: Josh
authors.initials: J
authors.lastname: Flook
authors.forename: Thomas
authors.initials: T
authors.lastname: Patterson
authors.forename: Kelly
authors.initials: K
authors.lastname: Robbins
authors.forename: Edward J
authors.initials: EJ
authors.lastname: Rebar
authors.forename: Philip D
authors.initials: PD
authors.lastname: Gregory
authors.forename: Fyodor D
authors.initials: FD
authors.lastname: Urnov
authors.forename: Joseph F
authors.initials: JF
authors.lastname: Petolino
mesh_terms: D015099:3-Oxoacyl-(Acyl-Carrier-Protein) Synthase / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D029688:Brassica napus / Q000201:enzymology* / Q000235:genetics*; D003433:Crosses, Genetic; D018076:DNA, Complementary / Q000235:genetics; D004789:Enzyme Activation; D005227:Fatty Acids / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D008969:Molecular Sequence Data; D018515:Plant Leaves / Q000201:enzymology / Q000235:genetics; D030821:Plants, Genetically Modified; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012639:Seeds / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D015533:Transcriptional Activation / Q000235:genetics*; D016335:Zinc Fingers / Q000235:genetics*
chemical_list: D018076:DNA, Complementary; D005227:Fatty Acids; D012333:RNA, Messenger; D014157:Transcription Factors; D015099:3-Oxoacyl-(Acyl-Carrier-Protein) Synthase