Engineered hypoxia biosensors and methods of using the same
AssigneeNorthwestern University
InventorsYannick Rene SCHREIBER, Joshua N. LEONARD, Patrick Sean DONAHUE
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Abstract
The present disclosure relates generally to hypoxia biosensors. In particular, the present disclosure relates to hypoxia biosensors that employ engineered genetic circuits for enhanced biosensor sensitivity and response. Some engineered genetic circuits employ positive feedback and amplification of expression in order to promote high-efficiency reporting of hypoxia.
Claims33
A DNA-based hypoxia biosensor, comprising:
- (a) a hypoxia-inducible promoter;
- (b) a nucleic acid sequence encoding a functional element;
- and
- (c) at least one nucleic acid sequence encoding a feedback element.
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A DNA-based hypoxia biosensor, comprising:
- (a) a hypoxia-inducible promoter;
- (b) a nucleic acid sequence encoding a functional element;
- (c) at least one nucleic acid sequence encoding one or more engineered proteins selected from the group consisting of:
- (i) an engineered protein that activates gene expression, wherein the engineered protein comprises a DNA binding domain and a transcription activator domain;
- (ii) an engineered protein that inhibits gene expression;
- (iii) an engineered protein that inhibits gene expression, the engineered protein optionally comprising one or more of a DNA binding domain, a bulky domain, a chromatin remodeling domain, and a transcription inhibitor domain;
- and
- (iv) a combination of two engineered proteins comprising a first engineered protein comprising a DNA binding domain fused to a dimerization domain, and a second engineered protein comprising a transcription regulator domain fused to a dimerization domain, wherein the dimerization domains of the two engineered proteins dimerize in the presence of a stimulus to which the dimerization domains of the two engineered proteins bind;
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- (d) one or more DNA binding sites for the DNA binding domain of the engineered protein(s) of (c).
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Dependencies are inferred from the claim text.