Screening for agents that target the actin cytoskeleton using c. elegans exposed to heat shock
AssigneeRutgers, The State University of New Jersey
InventorsFederico Sesti, Rahul Patel
Life of this patent
Estimating the term…
Abstract
Provided herein are screening methods that utilize transgenic nematodes exposed to heat shock conditions, such as agents that increase stability of F-actin. The transgenic nematodes used can be in a wild-type background, functionally deleted for OSG-1, or express human ARHGEF10 in an OSG-1 background. Such transgenic nematodes also express a fluorescent protein, such as GFP. Such methods are in some examples high throughput and automated. Also provided are recombinant nematodes and kits that can be used with such methods.
Claims24
A method for identifying an agent that targets actin, comprising:
- incubating in a culture medium a test agent with a transgenic nematode that expresses a fluorescent protein in a wild-type background;
- determining survival of the transgenic nematode prior to heat shock conditions;
- exposing the transgenic nematode to heat shock conditions;
- and
- determining survival rate for the transgenic nematode following the heat shock conditions, wherein a test agent that alters the ratio of survival of the transgenic nematode in the test agent after the heat shock conditions to survival of the transgenic nematode in the test agent before the heat shock conditions is an agent that targets actin.
A method for identifying an agent that targets actin, comprising:
- incubating a test agent with a transgenic first nematode that is functionally deleted for OSG-1 and expresses a fluorescent protein;
- incubating the test agent with a transgenic second nematode that expresses a functional mammalian ARHGEF10 protein and the fluorescent protein but does not express functional OSG-1;
- wherein the first and the second nematode are incubated separately in a culture medium,
- determining survival of the first and the second nematode prior to heat shock conditions;
- exposing the first and the second nematode to heat shock conditions;
- and
- determining survival rate for the first and the second nematode following the heat shock conditions, wherein a test agent that alters the ratio of survival of the transgenic second nematode in the test agent to survival of the transgenic first nematode in the test agent as compared to the ratio in an absence of the test agent is an agent that targets actin.
(canceled)
-8. (canceled)
(canceled)
(canceled)
An isolated transgenic nematode that expresses a functional mammalian ARHGEF10 protein and a fluorescent protein but does not express functional OSG-1.
Dependencies are inferred from the claim text.