Open AccessTranscriptional activation of Brassica napus β‐ketoacyl‐ACP synthase II with an engineered zinc finger protein transcription factor
Author(s) -
Gupta Manju,
DeKelver Russell C.,
Palta Asha,
Clifford Carla,
Gopalan Sunita,
Miller Jeffrey C.,
Novak Stephen,
Desloover Daniel,
Gachotte Daniel,
Connell James,
Flook Josh,
Patterson Thomas,
Robbins Kelly,
Rebar Edward J.,
Gregory Philip D.,
Urnov Fyodor D.,
Petolino Joseph F.
Publication year - 2012
Publication title -
plant biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.525
H-Index - 115
eISSN - 1467-7652
pISSN - 1467-7644
DOI - 10.1111/j.1467-7652.2012.00695.x
Subject(s) - canola , biology , zinc finger , transcription factor , gene , fatty acid synthase , brassica , transgene , transformation (genetics) , promoter , genetically modified crops , gene expression , genetics , botany
Summary 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.