Enhanced gene expression in algae
AssigneeSynaptic Research, LLC
InventorsGeorge Oyler, Julian Rosenberg
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Abstract
The invention provides a system of enhancing the expression of transgenes in algae. Transgenes are engineered to have a binding site for certain proteins in proximity to their promoter, for example a LexA binding site. The algae is also engineered to express a nucleosome alteration protein fused to a protein with affinity to the DNA binding site acting in coordination. An example is a LexA-p300 fusion protein, where the p300 is derived from Chlamydomonas. The LexA binding domain guides the p300 to the binding site and the p300 loosens the nucleosome structure by acetylating histones within proximity of the transgene, thus remodeling the local chromatin structure to allow for high-level expression.
Claims20
A construct for enhanced gene expression in an alga cell, comprising:
- a eukaryotic unicellular algae compatible transcriptional promoter functionally upstream of a coding sequence for a gene expression enhancer (GEE) fusion protein, wherein the fusion protein comprises an algae derived p300 functionally fused to the DNA binding protein, wherein at least the portion of the coding sequence of the DNA binding protein domain is codon optimized for improved expression in a eukaryotic unicellular algae;
- at least one transgene functionally downstream of a eukaryotic unicellular algae compatible transcriptional promoter;
- and
- at least one DNA region that is a binding site for the DNA binding protein, in vicinity of at least one of said transcriptional promoters.
A method of expressing a gene in an alga cell at higher levels, comprising:
- transforming eukaryotic unicellular algae with at least one vector comprising:
- a eukaryotic unicellular algae compatible transcriptional promoter functionally upstream of a coding sequence for a gene expression enhancer (GEE) fusion protein, wherein the fusion protein comprises an algae derived p300 functionally fused to the DNA binding protein, wherein at least the portion of the coding sequence of the DNA binding protein domain is codon optimized for improved expression in a eukaryotic unicellular algae;
- at least one transgene functionally downstream of a eukaryotic unicellular algae compatible transcriptional promoter;
- and
- at least one DNA region that is a binding site for the DNA binding protein, in vicinity of at least one of said transcriptional promoters;
- selecting a transformed algae cell;
- and
- detecting the expression of said GEE gene or said transgene in said algae.
Dependencies are inferred from the claim text.