Maize SPT Event Detection via Flanking Sequence Primers
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Solution Overview
Problem
Current methods for detecting specific transgenic events in plants are inefficient, as they require screening numerous events to identify optimal gene expression levels and patterns, and there is a need for reliable methods to detect the presence of transgenic sequences in progeny and products derived from recombinant crop plants for regulatory compliance and quality assurance.
Innovation Solution
Development of specific DNA primers and probes targeting the flanking sequences of the SPT event E6611.32.1.38 in maize, allowing for the identification of transgenic plants and products through nucleic acid amplification and hybridization techniques, enabling precise detection of the event in biological samples.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional screening methods are used to identify transgenic events with optimal gene expression, then the desired transgene expression levels and patterns can be achieved, but the process requires screening hundreds to thousands of events which is time-consuming and inefficient
Solution Approach 1:
The detection method is segmented into two distinct parts: a common primer that binds to conserved transgene sequences and an event-specific primer that binds to unique flanking sequences. This segmentation allows the method to efficiently distinguish between different transgenic events, enabling rapid identification of specific events without screening hundreds of candidates, thus resolving the contradiction between detection precision and screening efficiency.
Solution Approach 2:
The patent performs preliminary characterization of transgenic events by identifying and sequencing flanking sequences before developing detection methods. This preliminary action creates event-specific primers in advance, allowing for direct and efficient detection of specific transgenic events in regulatory and breeding contexts without requiring extensive screening, thereby improving productivity while maintaining detection precision.
2Reliability
If transgenic events are screened to ensure desired expression characteristics, then reliable gene expression can be achieved in adapted varieties, but the complexity of screening and characterizing numerous events increases
Solution Approach 1:
The method extracts and utilizes the unique flanking sequences that are specific to each transgenic event as the basis for detection. By taking out these event-specific sequences and designing primers based on them, the method simplifies the detection process to a single PCR reaction that directly identifies the specific event, reducing the complexity of screening multiple events while ensuring reliable detection of the desired transgene expression characteristics.
Solution Approach 2:
The detection method is designed with universal applicability through the common primer that can bind to various transgene constructs, while the event-specific primer provides specificity. This multi-functional design allows a single detection system to reliably identify multiple different transgenic events, maintaining reliability across diverse varieties and events without increasing procedural complexity.
3Measurement precision
If event-specific detection methods are developed to identify unique junctions between inserted DNA and recipient genome, then precise detection of specific transgenic events is enabled, but the requirement for characterizing and developing event-specific primers and probes increases the initial work required
Solution Approach 1:
The patent performs preliminary sequencing and characterization of flanking sequences during the transgenic event development phase. This preliminary action captures the event-specific genetic information early, which is then used to design specific primers and probes. By performing this characterization work in advance, the time required for event-specific detection is significantly reduced in subsequent regulatory and breeding applications, resolving the contradiction between detection precision and development time.
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
The solution enables reliable detection of the SPT event E6611.32.1.38 in plants and products, ensuring compliance with regulations and quality control, while facilitating marker-assisted breeding and segregation of transgenic material.
Implementation Method 1
Development of specific DNA primers and probes targeting the flanking sequences of the SPT event E6611.32.1.38 in maize, allowing for the identification of transgenic plants and products through nucleic acid amplification and hybridization techniques
Implementation Method 2
allowing for the identification of transgenic plants and products through nucleic acid amplification and hybridization techniques
Data Source
AI summary
Compositions and methods related to transgenic plants comprising seed production technology are provided. Specifically, maize plants having a E6611.32.1.38 event which confers seed production technology are provided. The plant harboring the E6611.32.1.38 event at the recited chromosomal location comprises the genomic/transgene junctions described. The plant genomic DNA flanking the integrated E6611.32.1.38 event can be used to design assays that will be specific for the E6611.32.1.38 event. The characterization of the genomic insertion site of the E6611.32.1.38 event provides for an enhanced breeding efficiency and enables the use of molecular markers to track the transgene insert in the breeding populations and progeny thereof. Various methods and compositions for the identification, detection, and use of the maize E6611.32.1.38 event are provided.