Cry Toxin Gene Mutations for Insecticidal Activity

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Solution Overview

Problem

There is a continuous need for new forms of pesticidal toxins to control insect pests effectively in agriculture, as existing methods may not fully address the devastation caused by insects and may have limitations in yield improvement and pest resistance.

Innovation Solution

The development of novel genes encoding pesticidal proteins, specifically nucleotide sequences and amino acid sequences, which can be used to confer pesticidal activity to bacteria, plants, and microorganisms, enabling the production of transgenic pest-resistant plants and formulations for controlling lepidopteran, hemipteran, coleopteran, nematode, and dipteran pests.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If existing pesticidal toxins are used to control insect pests, then some pest control is achieved, but the effectiveness is insufficient to fully address insect devastation and improve agricultural yields

Engineering Contradiction:
Improvepest control effectivenessVSAvoidagricultural yield improvement
Core Design Contradiction:
ReliabilityVSProductivity

Solution Approach 1:

The patent modifies existing Cry toxin genes through site-directed mutagenesis to create variant toxins with improved insecticidal activity. Specific amino acid substitutions are introduced to enhance binding affinity to insect gut receptors, thereby improving the reliability of pest control while addressing the insufficiency of existing toxins

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent creates hybrid toxin proteins by combining domains from different Cry toxin families (e.g., Cry1 and Cry2 domains) to generate novel pesticidal proteins with broader spectrum activity and enhanced effectiveness against multiple pest orders, thus improving both reliability and agricultural productivity

Inventive Principle:
Principle #40Composite materials

2Reliability

If novel pesticidal proteins are developed to enhance pest resistance, then pest control effectiveness improves, but the complexity of gene development and characterization increases

Engineering Contradiction:
Improvepest resistance effectivenessVSAvoidgene development complexity
Core Design Contradiction:
ReliabilityVSDevice complexity

Solution Approach 1:

The patent divides toxin proteins into functional domains (e.g., domain I for membrane insertion, domains II and III for receptor binding) that can be independently manipulated and recombined. This modular approach allows systematic development of variant toxins with reduced complexity compared to de novo protein design

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The patent develops a universal platform for toxin gene modification that can be applied across multiple Cry toxin families and pest targets. The same mutagenesis and domain recombination strategies are used to generate variants against different pest orders, reducing overall development complexity through method standardization

Inventive Principle:
Principle #6Universality (Multi-functionality)

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

These novel genes and proteins provide enhanced pest resistance and tolerance in plants, effectively killing or impairing pests, thereby improving agricultural yields and reducing pest-related damage.

Implementation Method 1

This toxin binds to apical brush border receptors in the midgut of the target larvae and inserts into the apical membrane creating ion channels or pores, resulting in larval death

Methodology Applied
Scientific EffectIon channel formation:

Implementation Method 2

The ingested protoxin is hydrolyzed by proteases in the insect digestive tract to an active toxic molecule

Methodology Applied
Scientific EffectHydrolysis: Hydrolysis

Data Source

PatentUS9862967B2AXMI281 toxin gene and methods for its use
Publication Date: 2018.01.09 BASF AGRICULTURAL SOLUTIONS US LLC

AI summary

Compositions and methods for conferring pesticidal activity to bacteria, plants, plant cells, tissues and seeds are provided. Compositions comprising a coding sequence for a toxin polypeptide are provided. The coding sequences can be used in DNA constructs or expression cassettes for transformation and expression in plants and bacteria. Compositions also comprise transformed bacteria, plants, plant cells, tissues, and seeds. In particular, isolated toxin nucleic acid molecules are provided. Additionally, amino acid sequences corresponding to the polynucleotides are encompassed, and antibodies specifically binding to those amino acid sequences. In particular, the present invention provides for isolated nucleic acid molecules comprising nucleotide sequences encoding the amino acid sequence shown in SEQ ID NO:2-5, or the nucleotide sequence set forth in SEQ ID NO: 1, as well as variants and fragments thereof.