AMPHIBIAN OOCYTE OR EMBRYO BIOASSAY
The present invention provides in vivo methods for screening compounds of interest. The methods rely on readily-observable phenotypic changes in amphibian oocytes or early embryos.
1 . An in vivo method for screening a plurality of compounds, the method comprising:
a) contacting a plurality of amphibian oocytes or embryos with the plurality of compounds; and
b) monitoring a phenotype in the plurality of amphibian oocytes or embryos to identify a compound that affects the phenotype.
2 . The method of claim 1 , wherein the method is performed in a multi-well format or a high throughput screening format.
3 . The method of claim 1 , wherein the monitoring step comprises time-lapse digital image capture.
4 . The method of claim 1 , wherein the phenotype is germinal vesicle breakdown, cell cleavage, a reporter-based assay, or an in-cell reporter assay.
5 . The method of claim 4 , wherein germinal vesicle breakdown is altered in a hormone-dependent manner or a hormone-independent manner; and cell cleavage is altered temporally, spatially, or both.
6 . The method of claim 1 , wherein the amphibian oocytes or embryos are from a Xenopus species or a Rana species.
7 . The method of claim 1 , wherein the amphibian oocytes or embryos are wild type, mutant, or genetically-modified.
8 . The method of claim 1 , wherein the plurality of compounds is a small molecule library, a pharmaceutically active compound library, a natural product library, a carbohydrate library, a lipid molecule library, a nucleic acid library, an antisense oligonucleotide library, a microRNA library, or a peptide library.
9 . The method of claim 1 , wherein the method further comprises c) determining a molecular mechanism of action for the compound.
10 . The method of claim 1 , wherein the plurality of compounds is a library of small molecules; the plurality of amphibian oocytes or embryos is from Xenopus laevis ; and the phenotype is germinal vesicle breakdown, cell cleavage, a reporter-based assay, or an in-cell reporter assay.
11 . A method for identifying a compound that affects a regulated mRNA translation control process, the method comprises:
a) contacting a plurality of amphibian oocytes or embryos with a plurality of compounds; and
b) monitoring a phenotype in the plurality of amphibian oocytes or embryos to identify the compound that affects the regulated mRNA translation control process.
12 . The method of claim 11 , wherein the method is performed in a multi-well format or a high throughput screening format.
13 . The method of claim 11 , wherein the monitoring step comprises time-lapse digital image capture.
14 . The method of claim 11 , wherein the phenotype is germinal vesicle breakdown, cell cleavage, a reporter-based assay, or an in-cell reporter assay.
15 . The method of claim 14 , wherein germinal vesicle breakdown is altered in a hormone-dependent manner or a hormone-independent manner; and cell cleavage is altered temporally, spatially, or both.
16 . The method of claim 11 , wherein the amphibian oocytes or embryos are from a Xenopus species or a Rana species.
17 . The method of claim 11 , wherein the amphibian oocytes or embryos are wild type, mutant, or genetically-modified.
18 . The method of claim 11 , wherein the plurality of compounds is a small molecule library, a pharmaceutically active compound library, a natural product library, a nucleic acid library, an antisense oligonucleotide library, a microRNA library, or a peptide library.
19 . The method of claim 11 , wherein the regulated mRNA translation process comprises a control protein chosen from Pumilio1, Pumilio2, Musashi1, Musashi2, or the cytoplasmic polyadenylation element binding protein (CPEB).
20 . The method of claim 11 , wherein the method further comprises c) determining a molecular mechanism of action for the compound.
21 . (canceled)