US 10,865,417 B2Granted

Yeast synthetic biology platform for identifying shikimate pathway enzyme inhibitors

AssigneesNew York University, Neochromosome Inc., The Johns Hopkins University

InventorsJef Boeke, Leslie Mitchell, Joe De Angelo, Joel S. Bader

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Abstract

Provided are compositions and methods for compound discovery. Modified yeast that have their endogenous yeast shikimate pathway disrupted or deleted, and replaced with homologous pathway genes from one or more distinct organisms, are provided and used in assays of test agents. The homologous pathway genes are designed to supplement the disrupted or deleted shikimate pathway genes. The assays are designed to identify whether or not the test agents can interfere with the function of enzymes in the shikimate pathway from organisms that are distinct from the yeast avatar hosts. In embodiments, the disruption/deletion of the yeast endogenous shikimate pathway results in the yeast being incapable of producing chorismic acid.

Claims7

Dependent claims

A method for identification of a test agent that can inhibit one or more enzymes that are part of a shikimate pathway, the method comprising introducing the test agent into modified Saccharomyces yeast, the modified Saccharomyces yeast comprising:

i) a disruption of an endogenous Saccharomyces yeast gene that encodes the shikimate pathway enzyme, and
ii) a supplemental gene encoding a heterologous enzyme that is homologous to the enzyme encoded by the endogenous Saccharomyces yeast gene;
wherein an inhibition of growth of the modified Saccharomyces yeast relative to a control is indicative that the test agent inhibits the shikimate pathway enzyme, and wherein the control comprises growth of the modified Saccharomyces yeast into which the test agent is introduced, and wherein the growth of the modified Saccharomyces yeast is performed on a medium that is supplemented with one or more aromatic amino acids such that growth of the modified yeast is independent of the function of the supplemental gene.

Dependencies are inferred from the claim text.

Facts

Application: 16/374,827Filed 4 Apr 2019 · us provisional application
Priority: 6 Apr 2018
Published: US 2019/0309314 A110 Oct 2019
Granted: 15 Dec 2020
Term ends: Not estimatedThe source records don’t support an estimate.

Classification16 codes

C12N 15/81
For fungi: for yeasts
C12N 1/185
Baker's yeast; Brewer's yeast: saccharomyces isolates
C12N 15/52
DNA or RNA fragments; Modified forms thereof ; Non-coding nucleic acids having a biological activity: genes encoding for enzymes or proenzymes
C12N 15/62
DNA or RNA fragments; Modified forms thereof ; Non-coding nucleic acids having a biological activity: dNA sequences coding for fusion proteins