	US 12,214,008 B2
	Protein interfaces
Charly Chahwan, San Francisco, CA (US); and Maria Soloveychik, San Francisco, CA (US)
Assigned to SyntheX, Inc., San Francisco, CA (US)
Filed by SyntheX, Inc., San Francisco, CA (US)
Filed on Feb. 9, 2021, as Appl. No. 17/171,841.
Application 17/171,841 is a continuation of application No. 16/215,432, filed on Dec. 10, 2018, abandoned.
Application 16/215,432 is a continuation of application No. 15/683,586, filed on Aug. 22, 2017, granted, now 10,188,691, issued on Jan. 29, 2019.
Application 15/683,586 is a continuation of application No. PCT/US2017/034870, filed on May 26, 2017.
Claims priority of provisional application 62/384,226, filed on Sep. 7, 2016.
Claims priority of provisional application 62/342,840, filed on May 27, 2016.
Prior Publication US 2021/0252100 A1, Aug. 19, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. A61K 38/02 (2006.01); A61K 38/00 (2006.01); A61K 39/00 (2006.01); A61K 45/06 (2006.01); C07K 5/11 (2006.01); C07K 7/06 (2006.01); C07K 7/08 (2006.01); C07K 14/00 (2006.01); C07K 16/30 (2006.01)

CPC A61K 38/02 (2013.01) [A61K 45/06 (2013.01); C07K 5/1019 (2013.01); C07K 7/06 (2013.01); C07K 7/08 (2013.01); C07K 14/00 (2013.01); C07K 16/30 (2013.01); A61K 38/00 (2013.01); A61K 2039/585 (2013.01); Y02A 50/30 (2018.01)]

8 Claims

1. A method for screening for inhibitors of a protein interaction between a first test protein and a second test protein, the method comprising:

expressing, in a plurality of host cells, a first fusion protein comprising a first test protein and a DNA-binding moiety; and

expressing, in each of the plurality of host cells, a second fusion protein comprising the second test protein and a gene-activating moiety,

wherein each of the plurality of host cells comprises a plurality of heterologous polynucleotide sequences that each encode a different cytotoxic agent, wherein expression of each of the heterologous polynucleotide sequences that each encode a different cytotoxic agent is under the control of a promoter that has selective affinity for the DNA-binding moiety, wherein interaction between the first test protein and the second test protein causes the gene-activating moiety to activate expression of each of the different cytotoxic agents encoded by each of the plurality of heterologous polynucleotide sequences;

wherein the DNA-binding moiety is a LexA, cI, Glucocorticoid receptor, TetR, or Ume6 DNA binding moiety;

wherein the gene activating moiety is a GAL4, B42, or Dof1 gene activating moiety;

wherein the plurality of different cytotoxic agents are selected from the group consisting of: a ribosomally-encoded xenobiotic agent, a ribosomally-encoded poison, a ribosomally-encoded recombinase that excises an essential gene for viability, and any combination thereof.