ATHB17 Leader Sequence Modulates Plant Gene Expression
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Solution Overview
Problem
In plant genetic engineering, achieving optimal gene expression in transgenic plants is challenging due to variables such as insertion site, regulatory sequence strength, codon usage, and presence of cryptic splice sites, often resulting in off-types and suboptimal phenotypes.
Innovation Solution
A recombinant polynucleotide comprising a genetic regulatory element, specifically the ATHB17-nt275-339 leader sequence, is operably linked to a transcribable polynucleotide, reducing transcription and translation levels, thereby improving phenotypes and avoiding off-types by acting as a leader for transcription and translation regulation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If strong regulatory sequences are used to increase gene expression, then productivity is improved, but manufacturing precision deteriorates due to off-types and suboptimal phenotypes
Solution Approach 1:
The patent modifies the regulatory sequence parameters by using the ATHB17-nt275-339 leader sequence, which has specific structural characteristics that modulate transcription and translation rates. This parameter change allows achieving desired expression levels while maintaining phenotype consistency and avoiding off-types.
Solution Approach 2:
The ATHB17-nt275-339 leader sequence acts as an intermediary element between the promoter and the coding region. It mediates the balance between achieving sufficient gene expression and maintaining precise phenotypic control, preventing both under-expression and over-expression related off-types.
2Productivity
If regulatory sequence strength is increased to improve gene expression, then productivity is improved, but reliability deteriorates due to variable expression patterns
Solution Approach 1:
The patent changes the regulatory sequence parameters by selecting the ATHB17-nt275-339 leader sequence with specific nucleotide composition and structure. This parameter optimization ensures stable and consistent gene expression patterns across different plants and tissues, improving reliability while maintaining productivity.
3Manufacturing precision
If multiple variables are adjusted to optimize transgene expression, then manufacturing precision is improved, but device complexity increases
Solution Approach 1:
The patent extracts and utilizes the specific functional elements present in the ATHB17-nt275-339 leader sequence that are responsible for optimal gene expression regulation. By focusing on this specific extracted sequence rather than adjusting multiple separate variables, the patent achieves expression optimization while minimizing construct complexity.
Data Source
AI summary
The disclosure provides DNA molecules and constructs, as well as their nucleotide sequences, useful for modulating gene expression in plants. The disclosure also provides nucleic acid sequences from Arabidopsis thaliana comprising novel regulatory elements identified from the AtHB17 gene. The disclosure also provides transgenic plants, plant cells, plant parts, and seeds comprising the DNA molecules operably linked to heterologous transcribable polynucleotides are also provided, as are methods of their use.