US 11,913,081 B2
Proximity-dependent split RNA polymerases as a versatile biosensor platform
Bryan C. Dickinson, Chicago, IL (US); Jinyue Pu, Chicago, IL (US); and Julia Zinkus, Chicago, IL (US)
Assigned to The University of Chicago, Chicago, IL (US)
Appl. No. 16/305,298
Filed by The University of Chicago, Chicago, IL (US)
PCT Filed Jun. 6, 2017, PCT No. PCT/IB2017/053324
§ 371(c)(1), (2) Date Nov. 28, 2018,
PCT Pub. No. WO2017/212400, PCT Pub. Date Dec. 14, 2017.
Claims priority of provisional application 62/424,875, filed on Nov. 21, 2016.
Claims priority of provisional application 62/346,174, filed on Jun. 6, 2016.
Prior Publication US 2020/0332371 A1, Oct. 22, 2020
Int. Cl. C12Q 1/6897 (2018.01); C12N 9/12 (2006.01); C12Q 1/689 (2018.01); C12Q 1/48 (2006.01)
CPC C12Q 1/6897 (2013.01) [C12N 9/1247 (2013.01); C12Q 1/48 (2013.01); C12Q 1/689 (2013.01); C12Y 207/07006 (2013.01)] 13 Claims
 
1. A system comprising:
a) a first polymerase domain attached to a first interaction component;
b) a second polymerase domain attached to a second interaction component;
c) a nucleic acid construct comprising a reporter gene under the control of a first heterologous promoter responsive to polymerase activity from the first and second polymerase domains when the first and second interaction components interact,
wherein the first polymerase domain comprises amino acids 1-179 of a T7 RNA polymerase and the second polymerase domain comprises amino acids 180-883 of a T7 RNA polymerase; wherein the system further comprises a negative control nucleic acid encoding a polypeptide that does not interact with the first or second interaction component; and wherein the polypeptide encoded by the negative control nucleic acid is attached to an alternate first polymerase domain comprising amino acids 1-179 of a T7 RNA polymerase or an alternate second polymerase domain comprising amino acids 180-883 of a T7 RNA polymerase and the system further comprises a second heterologous promoter responsive to polymerase activity from the interaction of the first and alternate second polymerase domains or the alternate first and the second polymerase domains.