Live yeast biosensors and methods of use thereof
AssigneeThe Trustees of Columbia University in the City of New York
InventorsVirginia Cornish, Tea CRNKOVIC, Davida RIOS, Amirhossein JAFARIYAN, Marco FANTINI, Anton KOZYRYEV
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Abstract
The present disclosure relates to kits, compositions and methods for detecting fungal species, viruses and/or protein variants in a sample, e.g., a biological sample. For example, but not by way of limitation, the present disclosure provides living yeast biosensors that have been genetically engineered to detect fungal species, viruses and/or protein variants in a sample, e.g., a biological sample.
Claims45
A genetically-engineered sensor cell for detecting the presence of a fungal species or a virus in a sample, comprising:
- (a) at least one heterologous G-protein coupled receptor (GPCR) that binds to an analyte derived from the fungal species or the virus, wherein the analyte is a ligand for the heterologous GPCR;
- and
- (b) a reporter gene, wherein binding of the analyte to the heterologous GPCR triggers the expression of the reporter gene and indicates the presence of the fungal species in the sample,
- wherein the fungal species is selected from the group consisting of A. nidulans, A. fumigatus, A. terreus, A. flavus, A. niger, A. clavatus, A. oryzae, A. novofumigatus, A. lentulus, A. viridimutans, A. udagawae, N. fischeri, T. citrinoviride, T. arundinaceum, T. longibrahiatum, T. harzianum, T. guizhouense, T. lentifore, T. virens, T. asperellum, T. gamsii, T. atroviride, B. dermatitidis, B. silverae, C. immitis, T. marnefei, P. carinii, P. murina, P. wakefieldiae, C. dubliniensis, C. auris, C. pseudohaemulonii, C. haemuloni, C. duobushaemulonis, C. metapsilosis, C. orthopsilosis and a combination thereof and/or wherein the virus is a respiratory virus, a hemorrhagic virus, a gastrointestinal virus, an exanthematous virus, a hepatitis virus, a sexually transmitted virus and/or a neurological virus.
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A method of detecting the presence of a fungal species or a virus in a sample, comprising:
- (a) contacting the sample with a sensor cell comprising at least one heterologous G-protein coupled receptor (GPCR) that binds to an analyte derived from the fungal species, wherein the analyte is a ligand for the heterologous GPCR;
- (b) binding of the analyte present in the sample to the heterologous GPCR, wherein binding of the analyte to the heterologous GPCR triggers an appearance of a reporter;
- and
- (c) detecting the appearance of the reporter, wherein the appearance of the reporter indicates the presence of the fungal species or the virus in the sample,
- wherein the fungal species is selected from the group consisting of A. nidulans, A. fumigatus, A. terreus, A. flavus, A. niger, A. clavatus, A. oryzae, A. novofumigatus, A. lentulus, A. viridinutans, A. udagawae, N. fischeri, T. citrinoviride, T. arundinaceum, T. longibrahiatum, T. harzianum, T. guizhouense, T. lentifore, T. virens, T. asperellum, T. gamsii, T. atroviride, B. dermatitidis, B. silverae, C. immitis, T. marnefei, P. carinii, P. murina, P. wakefieldiae, C. dubliniensis, C. auris, C. pseudohaemulonii, C. haemuloni, C. duobushaemulonis, C. metapsilosis, C. orthopsilosis and a combination thereof and/or wherein the virus is selected from the group consisting of a respiratory virus, a hemorrhagic virus, a gastrointestinal virus, an exanthematous virus, a hepatitis virus, a neurological virus, a sexually transmitted virus and a combination thereof.
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A genetically-engineered sensor cell for detecting the presence of a protein variant in a sample, comprising:
- (i) (a) at least one heterologous G-protein coupled receptor (GPCR) that binds to a wild type protein and one or more variants of the protein;
- (b) a protease that cleaves one or more of the protein variants;
- and
- (c) a reporter gene, wherein binding of the wild type protein to the heterologous GPCR triggers the expression of the reporter gene;
- or
- (ii) (a) at least one heterologous G-protein coupled receptor (GPCR) that binds to a wild type protein and one or more variants of the protein:
- (b) a protease that cleaves the wild type protein;
- and
- (c) a reporter gene, wherein binding of the one or more protein variants to the heterologous GPCR triggers the expression of the reporter gene.
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A genetically-engineered sensor cell for detecting an analyte in a sample, comprising:
- (a) a heterologous G-protein coupled receptor (GPCR) that binds to an analyte derived from the fungal species, wherein the analyte is a ligand for the heterologous GPCR;
- and
- (b) a reporter gene, wherein binding of the analyte to the heterologous GPCR triggers the expression of the reporter and indicates the presence of the fungal species in the sample, wherein the genetically-engineered sensor cell comprises a mutation and/or deletion in one or more of the following endogenous genes:
- Mfa1, Mfa2, Mf(alpha)1, Mf(alpha)2, Bar1, Far1, Sst2, Gpr1, Gpa2, Ste2, sfGFP, LEU2 and URA3.
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Dependencies are inferred from the claim text.