Maize Event DP-004114-3 Detection Using Segmented Primers
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for detecting the insect-resistant corn event DP-004114-3 are not discriminative enough, as they often rely on general genetic elements, making it difficult to distinguish between different events, especially those with similar DNA constructs, and there is a need for a simple and specific method to identify this event in biological samples for regulatory compliance and quality assurance.
Innovation Solution
A method using specific nucleic acid primers and probes that target unique sequences of the DP-004114-3 event, including the cry1F, cry34Ab1, cry35Ab1, and pat gene cassettes, to amplify or hybridize with DNA from the event, allowing for diagnostic amplicon production and detection, thereby identifying the event in corn samples.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If general genetic elements are used for detection, then detection broadness is improved, but measurement precision deteriorates
Solution Approach 1:
The detection system is segmented into multiple independent components: a first primer targeting a common genetic element present in all events of interest, and a second primer targeting an event-specific sequence. This segmentation allows the method to maintain broad detection capability while achieving high precision discrimination between different events.
Solution Approach 2:
The patent applies local quality by making the second primer event-specific rather than universal. Each event detection protocol uses a customized second primer that targets a unique sequence specific to that event, while the first primer remains general. This local customization at the primer design level enables precise discrimination without sacrificing overall detection broadness.
2Measurement precision
If event-specific primers are used, then measurement precision is improved, but device complexity increases
Solution Approach 1:
The patent merges the advantages of both general and specific detection approaches into a single unified PCR protocol. By combining a general first primer with an event-specific second primer in the same reaction mixture, the method achieves high precision event discrimination without requiring separate complex detection systems for each event.
Solution Approach 2:
The first primer serves multiple functions: it binds to a common genetic element shared by all events of interest, enabling universal detection across different events. This multi-functionality reduces overall system complexity by eliminating the need for separate universal amplification steps for each event.
3Adaptability or versatility
If multiple events are screened, then adaptability is improved, but loss of time increases
Solution Approach 1:
The patent enables periodic or sequential detection of multiple events using the same basic protocol framework. By maintaining a library of event-specific second primers, users can systematically screen for multiple events in a structured manner, reducing overall detection time compared to developing and validating separate detection methods for each event.
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 precise identification of DP-004114-3 in corn samples, ensuring compliance with regulations and quality standards by providing a discriminative and reliable method for detecting the event's presence, even in the presence of similar constructs.
Implementation Method 1
amplify or hybridize with DNA from the event, allowing for diagnostic amplicon production and detection
Implementation Method 2
specific nucleic acid primers and probes that target unique sequences of the DP-004114-3 event, including the cry1F, cry34Ab1, cry35Ab1, and pat gene cassettes, to amplify or hybridize with DNA from the event
Data Source
Figure 1
Figure 2
Figure 3
AI summary
The invention provides DNA compositions that relate to transgenic insect resistant maize plants. Also provided are assays for detecting the presence of the maize DP-004114-3 event based on the DNA sequence of the recombinant construct inserted into the maize genome and the DNA sequences flanking the insertion site. Kits and conditions useful in conducting the assays are provided.