US 12,065,479 B2
Force sensor cleavage domain containing chimeric polypeptides and methods of use thereof
Joseph H. Choe, San Francisco, CA (US); Paul Langridge, New York, NY (US); Wendell A. Lim, San Francisco, CA (US); Kole T. Roybal, San Francisco, CA (US); and Gary Struhl, New York, NY (US)
Assigned to THE REGENTS OF THE UNIVERSITY OF CALIFORNIA, Oakland, CA (US); and THE TRUSTEES OF COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK, New York, NY (US)
Appl. No. 16/763,524
Filed by The Regents of the University of California, Oakland, CA (US); and The Trustees of Columbia University in the City of New York, New York, NY (US)
PCT Filed Nov. 15, 2018, PCT No. PCT/US2018/061307
§ 371(c)(1), (2) Date May 12, 2020,
PCT Pub. No. WO2019/099689, PCT Pub. Date May 23, 2019.
Claims priority of provisional application 62/588,079, filed on Nov. 17, 2017.
Claims priority of provisional application 62/587,296, filed on Nov. 16, 2017.
Prior Publication US 2020/0331985 A1, Oct. 22, 2020
Int. Cl. C07K 14/755 (2006.01); C07K 14/59 (2006.01); C07K 14/705 (2006.01); C07K 14/72 (2006.01); C07K 16/28 (2006.01); G01N 33/50 (2006.01); G01N 33/68 (2006.01)
CPC C07K 14/755 (2013.01) [C07K 14/59 (2013.01); C07K 14/705 (2013.01); C07K 14/723 (2013.01); C07K 16/28 (2013.01); G01N 33/5005 (2013.01); G01N 33/6803 (2013.01); C07K 2317/54 (2013.01); C07K 2317/55 (2013.01); C07K 2317/622 (2013.01); C07K 2317/626 (2013.01); C07K 2319/02 (2013.01); C07K 2319/03 (2013.01); C07K 2319/50 (2013.01); C07K 2319/60 (2013.01)] 22 Claims
 
1. A chimeric polypeptide comprising, from N terminus to C terminus:
a) an extracellular domain comprising a first member of a binding pair;
b) a non-Notch force sensor cleavage domain comprising a proteolytic cleavage site;
c) a cleavable transmembrane domain; and
d) an intracellular domain that is not a Notch intracellular signaling domain and does not induce expression of Notch target genes,
wherein binding of the first member of the specific binding pair to the second member of the specific binding pair, present on a cell or other solid support, induces cleavage at the proteolytic cleavage site thereby releasing the intracellular domain, and
wherein the non-Notch force sensor cleavage domain is a von Willebrand Factor (vWF) A2 domain.