US 12,405,269 B2
Biosensors for measuring metastatic potential and chemoresistance of single cancer cells
Pradipta Ghosh, San Diego, CA (US)
Assigned to The Regents of the University of California, Oakland, CA (US)
Appl. No. 17/051,293
Filed by The Regents of the University of California, Oakland, CA (US)
PCT Filed May 13, 2019, PCT No. PCT/US2019/031954
§ 371(c)(1), (2) Date Oct. 28, 2020,
PCT Pub. No. WO2019/222071, PCT Pub. Date Nov. 21, 2019.
Claims priority of provisional application 62/670,905, filed on May 14, 2018.
Prior Publication US 2021/0239686 A1, Aug. 5, 2021
Int. Cl. G01N 33/542 (2006.01); G01N 33/574 (2006.01)
CPC G01N 33/542 (2013.01) [G01N 33/57496 (2013.01)] 6 Claims
 
1. A method for detecting metastatic potential and/or chemoresistance in single cells, comprising;
providing within a cell a multi-modular biosensor comprising:
only one stretch of the C-terminus of GIV flanking only one of tyrosines Y1764 and Y1798, wherein the stretch is capable of serving as a target substrate for one or more tyrosine kinases, wherein the stretch of the C-terminus of GIV flanking tyrosine Y1764 or Y1798 consists of 13 amino acids, and wherein the 13 amino acid sequence is selected from the group consisting of RKTEDTYFISSAG (SEQ ID No: 1) and SKDSNPYATLPRA (SEQ ID No: 2),
an N—SH2 domain of p85α (PI3K), and
a fluorescence resonance energy transfer (FRET) pair flanking the one or more stretches of the C-terminus of GIV and the N—SH2 domain of p85α (PI3K), the FRET pair comprising a donor fluorescent protein and an acceptor fluorescent protein; and
detecting FRET signal to measure metastatic potential.