US 12,410,487 B2
Selective modulation of protein-protein interactions
Maria Soloveychik, San Francisco, CA (US); and Charly Chahwan, San Francisco, CA (US)
Assigned to SyntheX, Inc., San Francisco, CA (US)
Filed by SyntheX, Inc., San Francisco, CA (US)
Filed on Nov. 1, 2023, as Appl. No. 18/499,563.
Application 18/499,563 is a continuation of application No. 15/931,295, filed on May 13, 2020, granted, now 11,840,743.
Application 15/931,295 is a continuation of application No. PCT/US2018/061292, filed on Nov. 15, 2018.
Claims priority of provisional application 62/590,147, filed on Nov. 22, 2017.
Claims priority of provisional application 62/587,269, filed on Nov. 16, 2017.
Prior Publication US 2024/0287626 A1, Aug. 29, 2024
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/80 (2006.01); C12N 15/10 (2006.01); C12N 15/81 (2006.01); C12Q 1/6897 (2018.01)
CPC C12Q 1/6897 (2013.01) [C12N 15/1055 (2013.01); C12N 15/1093 (2013.01); C12N 15/80 (2013.01); C12N 15/81 (2013.01); C12N 15/815 (2013.01); C12Q 2521/50 (2013.01); C12Q 2600/158 (2013.01)] 16 Claims
 
1. A method of screening for a molecule that disrupts an interaction between a first test protein and a second test protein in a host cell, while not disrupting an interaction between a third test protein and a second test protein in the host cell, the method comprising:
expressing in the host cell a first fusion protein comprising the first test protein and a first DNA-binding moiety;
expressing in the host cell a second fusion protein comprising the second test protein and a gene activating moiety;
expressing in the host cell a third fusion protein comprising a third test protein and a second DNA-binding moiety, wherein the second DNA-binding moiety is different from the first DNA-binding moiety; and
delivering a molecule to the host cell to test for disruption of the interaction of the first test protein and the second test protein without disrupting the interaction between the third test protein and the second protein;
wherein a plurality of death agent genes are disposed within the host cell, each operably linked to a promoter DNA sequence specific for the first DNA-binding moiety,
wherein a positive selection reporter is disposed within the host cell and operably linked to a promoter DNA sequence specific for the second DNA binding moiety,
wherein, when the molecule does not disrupt the interaction between the first test protein and the second test protein and does not disrupt the interaction between the second test protein and the third test protein, an interaction between the first test protein and the second test protein causes the gene activating moiety to activate expression of the death agent genes, while an interaction between the second test protein and the third test protein causes the gene activating moiety to activate expression of the positive selection reporter;
wherein, when the molecule disrupts the interaction between the first test protein and the second test protein but not the third test protein and the second test protein, (1) an interaction between the second test protein and the third test protein causes the gene activating moiety to activate expression of the positive selection reporter; and (2) disruption of the interaction between the first test protein and the second test moiety does not activate expression of the death agents, and
wherein the host cell is an animal cell.