Epigenetic Marker Identification via Yeast Histone Mutant Library
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Solution Overview
Problem
There is a need to elucidate the epigenetic commonalities between yeast sporulation and higher eukaryotic gametogenesis for studying mammalian gametogenesis and to identify epigenetic markers associated with mammalian infertility.
Innovation Solution
A yeast library comprising S. cerevisiae strains with histone mutations, where wild-type histone sequences are expressed from plasmids and substituted with mutant sequences, is used to identify epigenetic markers. The library is induced to sporulate, and mutants with diminished sporulation efficiency are assessed for epigenetic modifications, which are then correlated with mammalian gametogenesis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a yeast library with histone mutations is created to study epigenetic markers, then the ability to identify epigenetic markers is improved, but the complexity of the experimental system increases
Solution Approach 1:
The patent uses yeast (S. cerevisiae) as a simplified model system to copy and study epigenetic mechanisms that occur in mammalian gametogenesis. By creating a yeast library with histone mutations, the patent replicates key epigenetic processes in a more manageable organism, allowing identification of conserved epigenetic markers without the full complexity of mammalian systems.
Solution Approach 2:
The patent segments the histone protein into specific regions with defined mutations at particular amino acid positions. This segmentation allows systematic investigation of specific epigenetic modifications (such as phosphorylation at H3T11) while maintaining the overall functional integrity of the histone structure, thereby simplifying the analysis of epigenetic markers.
2Productivity
If histone sequences are mutated to create library strains, then the ability to assess sporulation efficiency is improved, but the difficulty of detecting and measuring epigenetic modifications increases
Solution Approach 1:
The patent introduces specific local mutations at defined amino acid positions within the histone sequences (such as mutating H3T11 to alanine). These localized changes create specific epigenetic markers that can be detected and measured, allowing assessment of sporulation efficiency while maintaining the ability to identify and quantify specific epigenetic modifications.
Solution Approach 2:
The patent employs detection methods that rely on identifying changes in the epigenetic state of histone modifications, analogous to detecting color changes. By creating histone mutants with specific modifications (such as phosphorylation sites), the patent enables visualization and measurement of epigenetic states through detectable changes that correlate with sporulation efficiency.
3Reliability
If wild-type histone sequences are expressed from plasmids, then the ability to control histone expression is improved, but the loss of time for library construction increases
Solution Approach 1:
The patent employs plasmid vectors that are pre-designed and pre-constructed with the necessary histone expression controls and mutation sites. By preparing these plasmids in advance with the correct sequences and regulatory elements, the patent enables rapid transformation into yeast strains without time-consuming library construction, while maintaining reliable control over histone expression.
Solution Approach 2:
The plasmid acts as an intermediary carrier that delivers the mutated histone sequences into the yeast genome. This intermediary system allows precise control over which histone variant is expressed and at what levels, while simplifying the overall process of creating the mutant library strains through a single transformation step rather than complex genetic manipulation.
Data Source
AI summary
The invention includes compositions comprising a S. cerevisiae yeast library, and methods of identifying an epigenetic marker for the diagnosis of infertility or a disorder associated with gametogenesis in an individual.