CRES-T Chimeric Repressor Peptides for Polyploid Gene Silencing
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Solution Overview
Problem
Conventional methods for repressing gene expression in plants, such as antisense and ribozyme techniques, face challenges in designing specific sequences for practical plants due to genetic variability and redundancy, making it difficult to disrupt target genes effectively, especially in polyploid organisms like soybean and wheat.
Innovation Solution
Development of a chimeric repressor silencing technology (CRES-T) using a conserved transcriptional repression domain (L/F)DLN(L/F)(X)P, which is fused with a transcriptional activator to create a chimeric protein that represses transcription of target genes and related redundant genes, eliminating the need for sequence-specific design.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional antisense or ribozyme methods are used to repress target gene expression, then sequence-specific repression can be achieved, but the complexity of designing and implementing sequence-specific sequences increases due to genetic variability and redundancy in practical plants
Solution Approach 1:
The patent applies universality by using a transcriptional repressor peptide with a conserved motif that can repress multiple target genes with different sequences. The repressor peptide contains a conserved motif (e.g., RXLFGVNM) that enables it to bind to and repress transcription of various target genes, eliminating the need for gene-specific sequence design. This allows a single repressor construct to function across multiple genes and plant types.
Solution Approach 2:
The patent applies parameter changes by focusing on conserved motifs rather than variable sequences. Instead of designing sequences based on specific gene sequences that vary between plants, the invention identifies and utilizes conserved motifs (such as RXLFGVNM) that remain unchanged across different genes and plant species. This parameter shift from sequence-specific to motif-based design simplifies the approach while maintaining repression effectiveness.
2Reliability
If gene knock-out method is used to disrupt target genes, then complete gene disruption can be achieved, but the difficulty increases significantly in polyploid plants with redundant gene sets
Solution Approach 1:
The patent applies local quality by using a transcriptional repressor that specifically targets and represses transcription of particular genes through binding to their promoters. Rather than attempting to disrupt all genes in a polyploid genome, the repressor is designed to locally repress specific target genes with the conserved motif. This allows selective repression of individual genes or gene families without affecting other genes, making the process manageable even in complex polyploid genomes.
Solution Approach 2:
The patent uses a transcriptional repressor peptide as an intermediary mechanism between the introduced construct and the target genes. The repressor peptide acts as a mediator that binds to promoter regions of target genes and blocks transcription initiation. This intermediary approach avoids the need for direct gene disruption through chemical or physical means, providing a more controllable and implementable method for gene silencing in polyploid plants.
3Measurement precision
If model plants are used for genetic studies, then well-understood gene sequences are available, but the applicability to practical plants with different gene sequences is limited
Solution Approach 1:
The patent applies universality by developing a transcriptional repressor system that functions across different plant types. The conserved motif in the repressor peptide allows it to recognize and repress target genes in both model plants and practical crops. This universal repressor construct can be applied to various plant species without requiring species-specific design, thereby bridging the gap between model plant research and practical plant improvement.
Solution Approach 2:
The patent applies parameter changes by shifting the design criterion from model-specific sequences to conserved motifs that are preserved across plant species. By focusing on conserved elements (such as the RXLFGVNM motif) rather than variable sequences, the repressor system maintains its effectiveness across different plant types. This parameter change enables the same repressor construct to work in both model plants and practical crops with different gene sequences.
Data Source
AI summary
Provided is a peptide which is capable of repressing transcription in a plant and which can be used in the CRES-T (a simple and widely applicable means for repressing gene transcription). Also provided is a gene encoding the peptide.