Chimeric Promoter Regulatory Elements for Root-Preferred Transgene Expression

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Solution Overview

Problem

Current plant transformation technologies face challenges in achieving high expression of multiple transgenes at a single genomic locus and tissue-specific expression of transgenes, leading to inefficiencies in conferring desirable traits such as pest resistance and herbicide tolerance.

Innovation Solution

The development of chimeric promoter regulatory elements, specifically combining upstream promoters, introns, and 5′-UTRs, to create a novel gene expression cassette that enhances transgene expression in plant cells and tissues, including root-preferred expression.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Quantity of substance

If conventional plant transformation technologies are used to introduce multiple transgenes, then the number of transgenes can be increased, but the expression level of each transgene decreases

Engineering Contradiction:

Improvenumber of transgenesVSAvoidexpression level

Core Design Contradiction:

Quantity of substanceVSReliability

Solution Approach 1:

The patent combines multiple regulatory elements (promoter, intron, 5'-UTR) from different sources into a single chimeric regulatory element that can drive high-level expression of multiple transgenes simultaneously, resolving the contradiction between transgene number and expression level

Inventive Principle:

Principle #5Merging (Combining)

Solution Approach 2:

The chimeric regulatory element is constructed as a composite of multiple functional DNA sequences (promoter region from Zea mays Peroxidase 5, intron from Zea mays alcohol dehydrogenase, 5'-UTR from Maize Streak Virus), creating a superior regulatory element that achieves both high expression and multi-transgene compatibility

Inventive Principle:

Principle #40Composite materials

2Adaptability or versatility

If conventional promoters are used for transgene expression, then general expression can be achieved, but tissue-specific expression cannot be obtained

Engineering Contradiction:

Improveexpression flexibilityVSAvoidtissue-specificity

Core Design Contradiction:

Adaptability or versatilityVSManufacturing precision

Solution Approach 1:

The chimeric regulatory element incorporates the Maize Streak Virus 5'-UTR which contains specific sequences that direct expression to particular plant tissues or developmental stages, enabling spatial and temporal control of transgene expression while maintaining high expression levels

Inventive Principle:

Principle #3Local quality

Data Source

PatentUS9909133B2Root specific expression conferred by chimeric gene regulatory elements

Publication Date: 2018.03.06 CORTEVA AGRISCIENCE LLC

AI summary

Provided are constructs and methods for expressing a transgene in plant cells and/or plant tissues using chimeric gene regulatory elements.