Labyrinthulomycetes Regulatory Elements for Transgene Expression Control
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Solution Overview
Problem
Current genetic modification techniques face challenges in achieving optimal and temporally specific expression of transgenes in recombinant cells, particularly in microbial organisms, due to insufficient expression levels and nonspecific expression patterns, necessitating the development of novel regulatory elements that can control gene expression across different growth phases and conditions.
Innovation Solution
The use of isolated, synthetic, or recombinant nucleic acid molecules with nucleic acid sequences exhibiting high stringency hybridization or sequence identity to specific sequences (SEQ ID NOs:1-70 and 180-202) that function as promoters or regulatory elements, operably linked to heterologous nucleic acid sequences, to drive gene expression in labyrinthulomycetes cells under various conditions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional regulatory elements are used to control transgene expression, then gene expression can be achieved, but the expression level is insufficient and lacks temporal specificity
Solution Approach 1:
The patent applies parameter changes by utilizing promoters with varying strengths (weak, moderate, strong) to precisely control the level and timing of transgene expression. Different promoter strengths enable expression to be tuned according to specific developmental stages or physiological conditions, thereby achieving both high expression levels when needed and temporal specificity by selecting appropriate promoter strength for each application context.
2Device complexity
If the same regulatory sequence is used to control multiple transgenes, then device complexity is reduced, but undesirable molecular interactions occur
Solution Approach 1:
The patent provides a universal collection of regulatory sequences from labyrinthulomycetes that can be applied to control multiple different transgenes. Each regulatory sequence in the collection serves multiple potential functions across different genetic engineering applications, allowing researchers to select appropriate sequences for controlling expression of various transgenes without needing to develop new regulatory elements for each case, thus reducing overall system complexity while maintaining reliability through sequence diversity.
3Adaptability or versatility
If genetic modification techniques are applied to achieve desirable traits, then commercially desirable characteristics are obtained, but expression control precision is insufficient
Solution Approach 1:
The patent segments the gene expression control system into distinct functional components: different promoter types (weak, moderate, strong), various regulatory sequences, and specific labyrinthulomycetes-derived elements. This segmentation allows precise control of each transgene's expression level and timing independently, enabling multiple desirable traits to be introduced into recombinant organisms with precise expression control for each trait, thereby achieving both trait diversity and manufacturing precision.
Data Source
AI summary
The present disclosure generally relates to novel polynucleotide molecules for use in regulating gene expression in recombinant cells, such as labyrinthulomycetes cells. The disclosure further relates to nucleic acid constructs, such as vectors and expression cassettes, containing a regulatory element operably linked to a heterologous nucleotide sequence. The disclosure further relates to methods for stably transforming a host cell, such as a labyrinthulomycetes cell with transgenes. Stably transformed recombinant cells, progeny, biomaterials derived therefrom, and methods for preparing and using the same are also provided.