Methods for assessing the delivery of exogenous agents
AssigneeAbbott Laboratories
InventorsYu Shen, Xiaoyu Lin, Stephen W. Fesik, Leiming Li
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Abstract
This invention provides methods, compositions and kits for rapid determination of the delivery of exogenous agents both in vitro and in vivo, including without limitation siRNA, microRNA, a ribozyme or an antisence molecule, any of which may target, bind to, or inactivate the mRNA of the gene of interest expressed in the cells. The methods, compositions and kits utilize a promoter-reporter construct whereby successful non-viral nucleic acid delivery leads to an up-regulation of reporter signals thus providing a quantitative, sensitive and rapid means of detection, validation and monitoring.
Claims40
A method of analyzing the delivery of an exogenous agent to a tissue comprising:
- (a) providing a tissue that expresses a fusion protein, wherein said fusion protein has a half-life of no more than about 72 hours;
- (b) contacting said tissue with the exogenous agent;
- (c) analyzing a sample from the tissue, thereby providing analysis of the delivery of the exogenous agent to the tissue.
A method of analyzing the delivery of an exogenous agent to a tissue comprising:
- (a) providing a tissue having the tet fusion protein which further comprises a PEST sequence, wherein said PEST sequence is a PEST sequence-containing portion of a C-terminus of murine ornithine decarboxylase (MODC);
- (b) contacting said tissue with the agent;
- (c) analyzing a sample from the tissue, thereby providing analysis of the delivery of the exogenous agent to the tissue.
A transgenic non-human animal comprising a recombinant nucleic acid molecule stably integrated into the genome of said animal, said molecule encoding a tet repressor fusion protein further comprises a PEST sequence, wherein said PEST sequence is a PEST sequence-containing portion of a C-terminus of murine ornithine decarboxylase (MODC).
A method of analyzing the delivery of an exogenous agent to a tissue comprising:
- (a) providing a non-human transgenic non-human animal comprising a recombinant nucleic acid molecule stably integrated into the genome of said animal, said molecule encoding a tet fusion protein further comprises a PEST sequence, wherein said PEST sequence is a PEST sequence-containing portion of a C-terminus of murine ornithine decarboxylase (MODC);
- (b) contacting said non-human animal with the agent;
- (c) analyzing a sample from the tissue of the non-human animal, thereby providing analysis of the delivery of the exogenous agent to the tissue.
A method of analyzing the delivery of an exogenous agent to a tissue comprising:
- (a) providing a non-human transgenic non-human animal comprising a recombinant nucleic acid molecule stably integrated into the genome of said animal, said molecule encoding a tet fusion protein further comprises a PEST sequence, wherein said PEST sequence is a PEST sequence-containing portion of a C-terminus of murine ornithine decarboxylase (MODC);
- (b) contacting said non-human animal with the agent;
- (c) non-invasively detecting the delivery of the exogenous agent to the tissue.
The method of claim 39, wherein said imaging is bioluminescence imaging.
Dependencies are inferred from the claim text.