Multi-Promoter Transgenic Event for Selective Gene Expression

Solution Overview

Problem

Current methods for transgenic plant production require costly and time-consuming re-cloning and production of additional transgenic events to achieve altered or specific expression patterns of genes of interest, limiting the efficiency of gene function studies and plant breeding strategies.

Innovation Solution

The method involves genetic crosses between parent plants with a target RNA and those having expression-specific promoters for suppressing the target RNA, allowing for selective and rapid alteration of transgene expression patterns in a single generation, using DNA that transcribes to single-stranded or double-stranded RNA for silencing.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Manufacturing precision

If re-cloning and production of additional transgenic events is performed to achieve altered expression patterns, then specific expression patterns can be obtained, but time and cost increase significantly

Engineering Contradiction:

Improveexpression pattern specificityVSAvoidtime for generating transgenic events

Core Design Contradiction:

Manufacturing precisionVSLoss of time

Solution Approach 1:

The patent applies preliminary action by pre-integrating multiple different promoter sequences into a single transgenic event construct before transformation. This allows all promoter variants to be available in advance within one plant genome, eliminating the need to perform separate re-cloning and transformation procedures for each promoter variant, thus significantly reducing the time required while maintaining the ability to achieve specific expression patterns.

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The patent implements universality by designing a multi-functional transgenic event that contains multiple promoter sequences (e.g., promoter A, promoter B, promoter C) within a single construct. This single transgenic event can produce multiple expression patterns by alternatively transcribing from different promoters, replacing the need for multiple separate transgenic events, each dedicated to a single promoter variant, thereby reducing both time and resource requirements.

Inventive Principle:

Principle #6Universality (Multi-functionality)

2Manufacturing precision

If re-cloning and production of additional transgenic events is performed to achieve altered expression patterns, then specific expression patterns can be obtained, but cost increases significantly

Engineering Contradiction:

Improveexpression pattern specificityVSAvoidcost of generating transgenic events

Core Design Contradiction:

Manufacturing precisionVSEase of manufacture

Solution Approach 1:

The patent implements universality by designing a multi-functional transgenic event that contains multiple promoter sequences (e.g., promoter A, promoter B, promoter C) within a single construct. This single transgenic event can produce multiple expression patterns by alternatively transcribing from different promoters, replacing the need for multiple separate transgenic events, each dedicated to a single promoter variant, thereby reducing both time and resource requirements.

Inventive Principle:

Principle #6Universality (Multi-functionality)

Solution Approach 2:

The patent applies merging by combining multiple promoter sequences into a single integrated transgenic construct. Instead of creating separate transgenic events for each promoter, the invention merges them into one event that can be transformed into the plant genome in a single procedure, reducing the cumulative cost of multiple transformation experiments while maintaining the capability to generate specific expression patterns.

Inventive Principle:

Principle #5Merging (Combining)

3Loss of information

If multiple transgenic events are produced to assess promoter effects, then comprehensive data can be obtained, but the process becomes complex and time-consuming

Engineering Contradiction:

Improvecompleteness of promoter effect dataVSAvoidcomplexity of transgenic event production

Core Design Contradiction:

Loss of informationVSDevice complexity

Solution Approach 1:

The patent applies preliminary action by pre-integrating multiple different promoter sequences into a single transgenic event construct before transformation. This allows all promoter variants to be available in advance within one plant genome, eliminating the need to perform separate re-cloning and transformation procedures for each promoter variant, thus significantly reducing the time required while maintaining the ability to achieve specific expression patterns.

Inventive Principle:

Principle #10Preliminary action

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

This approach enables convenient and rapid assessment of promoter effects on gene expression, optimizing plant breeding by allowing selective expression of target RNAs in specific tissues, thereby reducing the time and cost associated with generating desired phenotypes and improving the understanding of gene function.

Implementation Method 1

an expression-specific promoter operably linked to DNA for suppressing expression of the target RNA

Methodology Applied

Scientific EffectGene silencing:

Data Source

PatentUS9121028B2Selective gene expression in plants

Publication Date: 2015.09.01 MONSANTO TECHNOLOGY LLC

AI summary

This invention discloses methods for providing plants with selective expression of a target RNA and methods for evaluating an expression pattern of a target RNA in a plant. Also disclosed are transgenic plants having recombinant DNA for expressing a protein using a promoter functional in multiple tissues, and recombinant DNA for suppressing expression of the protein using a promoter functional in fewer than the multiple tissues.