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US 2022/0119825 A1Published application
Scalable peptide-gpcr intercellular signaling systems
AssigneeThe Trustees of Columbia University in the City of New York
InventorsVirginia Cornish, James Brisbois, Sonja Billerbeck, Miguel Jimenez
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Abstract
The present disclosure relates to intercellular signaling between genetically-engineered cells and, more specifically, to a scalable peptide-GPCR intercellular signaling system. The present disclosure provides an intercellular signaling system that includes at least two cells that have been genetically-engineered to communicate with each other, methods of use and kits thereof.
Claims20
A genetically-engineered cell expressing:
- (a) at least one heterologous G-protein coupled receptor (GPCR), wherein the amino acid sequence of the heterologous GPCR is at least about 75% homologous to an amino acid sequence comprising any one of SEQ ID NOs:
- 117-161 or an amino acid sequence provided in Table 11 and/or is encoded by a nucleotide sequence that is at least about 75% homologous to a nucleotide sequence comprising any one of SEQ ID NOs:
- 168-211;
- and/or
- (b) at least one heterologous secretable GPCR peptide ligand, wherein the amino acid sequence of the heterologous GPCR peptide ligand is at least about 75% homologous to an amino acid sequence comprising any one of SEQ ID NOs:
- 1-116 or an amino acid sequence provided in Table 12 and/or encoded by a nucleotide sequence that is about 75% homologous to a nucleotide sequence comprising any one of SEQ ID NOs:
- 215-230.
4 more elements
An intercellular signaling system comprising:
- (i) (a) a first genetically-engineered cell expressing at least one secretable G-protein coupled receptor (GPCR) ligand;
- and (b) a second genetically-engineered cell expressing at least one heterologous GPCR, wherein (i) the amino acid sequence of the heterologous GPCR is at least about 75% homologous to an amino acid sequence comprising any one of SEQ ID NOs:
- 117-161 or an amino acid sequence provided in Table 11 and/or is encoded by a nucleotide sequence that is at least about 75% homologous to a nucleotide sequence comprising any one of SEQ ID NOs:
- 168-211 and/or (ii) the amino acid sequence of the secretable GPCR ligand is at least about 75% homologous to an amino acid sequence comprising any one of SEQ ID NOs:
- 1-116 or an amino acid sequence provided in Table 12 and/or is encoded by a nucleotide sequence that is about 75% homologous to a nucleotide sequence comprising any one of SEQ ID NOs:
- 215-230;
- or
- (ii) (a) a first genetically-engineered cell comprising:
- (i) a nucleic acid encoding a first heterologous G-protein coupled receptor (GPCR);
- and/or (ii) a nucleic acid encoding a first secretable GPCR ligand;
- and (b) a second genetically-engineered cell comprising:
- (i) a nucleic acid encoding a second heterologous GPCR;
- and/or (ii) a nucleic acid encoding a second secretable GPCR ligand, wherein (i) the first GPCR and/or the second GPCR is at least about 75% homologous to an amino acid sequence comprising any one of SEQ ID NOs:
- 117-161 or an amino acid sequence provided in Table 11 and/or is encoded by a nucleotide sequence that is at least about 75% homologous to a nucleotide sequence comprising any one of SEQ ID NOs:
- 168-211;
- and/or (ii) the first and/or second secretable GPCR peptide ligand is at least about 75% homologous to an amino acid sequence comprising any one of SEQ ID NOs:
- 1-116 or an amino acid sequence provided in Table 12 and/or is encoded by a nucleotide sequence that is about 75% homologous to a nucleotide sequence comprising any one of SEQ ID NOs:
- 215-230.
15 more elements
A method for the identification of a G-protein coupled receptor (GPCR) and/or a GPCR ligand to be expressed in a genetically-engineered cell, comprising:
- (a) searching a protein and/or genomic database and/or literature for a protein and/or a gene with homology to:
- (i) a S. cerevisiae Ste2 receptor and/or Ste3 receptor;
- (ii) a GPCR comprising an amino acid sequence comprising any one of SEQ ID NOs:
- 117-161;
- (iii) a GPCR comprising an amino acid sequence provided in Table 11;
- and/or (iv) a GPCR encoded by a nucleotide sequence comprising any one of SEQ ID NOs:
- 168-211 to identify a GPCR;
- and/or
- (b) searching a protein and/or genomic database and/or literature for a protein, peptide and/or a gene with homology to:
- (i) a GPCR peptide ligand comprising an amino acid sequence comprising any one of SEQ ID NOs:
- 1-116;
- (ii) a GPCR peptide ligand comprising an amino acid sequence provided in Table 12;
- (iii) a GPCR peptide ligand encoded by a nucleotide sequence comprising any one of SEQ ID NOs:
- 215-230 to identify a GPCR ligand;
- and/or (iv) a yeast pheromone or a motif thereof.
12 more elements
Dependencies are inferred from the claim text.