High performance multi-input microRNA sensors and uses thereof
AssigneeMassachusetts Institute of Technology
InventorsRon Weiss, Jin Huh
Life of this patent
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Abstract
Provided herein are genetic circuits and cell state classifiers for detecting the microRNA profile of a cell. The cell state classifiers of the present disclosure are designed to incorporate multiple genetic circuits integrated together by transcriptional or translational control. Multiple inputs can be sensed simultaneously by coupling their detection to different portions of the genetic circuit such that the output molecule is produced only when the correct input profile of miRNAs is detected. The genetic circuits and cell state classifiers may be used in various applications (e.g., therapeutic or diagnostic applications).
Claims17
A cell state classifier, the cell state classifier comprising:
- (i) a first sensor circuit comprising a promoter operably linked to a nucleotide sequence encoding a first activator and one or more target sites for a first set of microRNAs;
- (ii) a second sensor circuit comprising:
- (a) a promoter operably linked to a nucleotide sequence encoding a second activator, one or more target sites for a second microRNA, and one or more target sites for a first regulatory microRNA that is different from any of the first set of microRNAs or the second microRNA;
- (b) a promoter that is activated by the second activator of (ii)(a), operably linked to a nucleotide sequence encoding a first repressor and one or more target sites for the second microRNA of (ii)(a);
- wherein the second activator is different from the first activator,
- (iii) a first signal circuit comprising:
- (a) a first activatable/repressible promoter that is activated by the first activator of (i) or repressed by the first repressor of (ii)(b), operably linked to a nucleotide sequence encoding a first output molecule, to a nucleotide sequence encoding a third activator, and to a nucleotide sequence encoding the first regulatory microRNA;
- and
- (b) one or more target sites for any one of the first set of microRNAs of (i);
- and
- (iv) a second signal circuit comprising:
- (a) a promoter that is activated by the third activator of (iii)(a) in the presence of an inducer, operably linked to a nucleotide sequence encoding a second output molecule, and
- (b) one or more target sites for a second regulatory microRNA that is different from any of the first set of microRNAs, the second microRNA, or the first regulatory microRNA.
Dependencies are inferred from the claim text.