Corn Elite Event MZIR260 Junction Detection for Event Discrimination
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Solution Overview
Problem
Existing methods for detecting and distinguishing between different transgenic events in plants expressing Cry proteins are inadequate, particularly for high-quality events like MZIR260, due to reliance on interchangeable coding sequences and lack of specificity in current nucleic acid detection methods, which fail to discriminate between events with similar constructs.
Innovation Solution
The development of novel nucleic acid sequences, including junction sequences unique to elite event MZIR260, allows for specific detection of MZIR260 through the use of polynucleotide primers and probes, enabling accurate identification of the event in biological samples.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional nucleic acid detection methods are used to detect transgenic events, then detection can be performed, but the methods lack specificity and fail to discriminate between events with similar constructs
Solution Approach 1:
The patent segments the detection approach by developing event-specific junction sequences that distinguish MZIR260 from other transgenic events. Instead of using generic detection methods, the invention divides the detection space into event-specific segments through unique primer and probe designs targeting the specific genomic insertion site of MZIR260, thereby achieving high discrimination capability.
Solution Approach 2:
The patent introduces junction sequences as intermediary elements that bridge the gap between generic transgenic detection and event-specific identification. These junction sequences serve as unique mediators that allow detection methods to specifically recognize MZIR260 by targeting the unique genomic context surrounding the transgene insertion, rather than detecting the transgene itself which is common across multiple events.
2Reliability
If hundreds of different events are produced and screened to find high-quality events, then elite events with desired molecular qualities can be identified, but the process is time-consuming and resource-intensive
Solution Approach 1:
The patent applies preliminary action by pre-characterizing the genomic insertion sites and developing detection methods for elite events like MZIR260 during the event development phase. This allows breeders and regulators to have ready-to-use, validated detection tools before commercial deployment, eliminating the need for time-consuming screening processes when the events are introduced to the market.
Solution Approach 2:
The patent uses copying by creating reproducible detection protocols based on the characterized junction sequences of elite events. Once the unique genomic context of MZIR260 is identified and validated, the detection method can be copied and standardized across laboratories and regulatory bodies, ensuring consistent identification without requiring re-screening.
3Object-affected harmful factors
If transgenic events are introduced into commercial cultivation, then insect resistance can be provided, but resistance breakdown occurs over time due to field-evolved insect resistance
Solution Approach 1:
The patent applies parameter changes by introducing novel Cry proteins with different amino acid sequences and structural characteristics (e.g., eCry3.1Ab, Cry34Ab1, Cry35Ab1) that target insect pests through different mechanisms. These parameter changes in protein structure and mode of action create new resistance barriers that are effective against pests that have evolved resistance to previous Cry proteins.
4Ease of operation
If detection methods focus on frequently used genetic elements like promoters and terminators, then simplicity and uniformity of reagents are achieved, but the methods cannot discriminate between transgenic constructs that differ only in coding sequences
Solution Approach 1:
The patent applies local quality by shifting the detection focus from universal genetic elements (promoters, terminators) to the local, event-specific genomic context at the insertion site. The junction sequences represent the unique local quality of each transgenic event's integration into the corn genome, allowing detection methods to maintain simplicity while achieving event-specific discrimination through targeted sequencing or PCR approaches.
Data Source
AI summary
A novel transgenic corn elite event designated MZIR260 is disclosed. The invention relates to DNA sequences of the recombinant constructs inserted into the corn genome and of genomic sequences flanking the insertion site that resulted in elite event MZIR260. The invention further relates to assays for detecting the presence of the DNA sequences of corn elite event MZIR260, to corn plants and corn seeds comprising the genotype thereof, and to methods for producing a corn plant by crossing a corn plant comprising the elite event MZIR260 genotype with itself or another corn variety.