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US 10,378,070 B2Granted
Vector and gene introduction agent for monitoring and visualizing cell differentiation using expression of micro RNA as indicator and monitoring and visualizing method using thereof
AssigneeKyoto University
InventorsHirohide Saito, Hideyuki Nakanishi
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Abstract
Provided is a method for continuously visualizing and accurately determining a living cellular state per se. Disclosed are: a vector comprising 5′- and 3′-end nucleic acid sequences for integration and a reporter gene sequence positioned between the 5′- and 3′-end nucleic acid sequences for integration; a gene introduction agent comprising a first vector and a second vector; a cell comprising the vector or the agent; and a method for determining a cellular state by using the cell.
Claims7
A vector comprising:
- (I) 5′- and 3′-end nucleic acid sequences for integration to be recognized by a transposase, and
- (II) a reporter gene sequence positioned between the 5′- and 3′-end nucleic acid sequences for integration, the reporter gene sequence comprising:
- in this order in the 5′ to 3′ direction, a) an inducible promoter;
- b) a sequence encoding a first marker protein;
- c) a poly(A) substitute sequence;
- d) an RNaseP/Z cleavage site sequence;
- e) an mRNA-stabilizing sequence;
- f) a sequence to drive polycistronic expression;
- g) an miRNA target sequence;
- h) a sequence encoding a second marker protein;
- i) a poly(A) substitute sequence and an RNaseP/Z cleavage site sequence, or a poly(A) signal sequence;
- and j) a sequence encoding an activator for the inducible promoter.
9 more elements
Dependencies are inferred from the claim text.