US 11,656,221 B2
Methods to identify modulators of actin-binding proteins
Adam William Avery, Rochester Hills, MI (US); Thomas S. Hays, St. Paul, MN (US); David D. Thomas, Minneapolis, MN (US); Michael E. Fealey, Dallas, TX (US); and Robyn T. Rebbeck, Minneapolis, MN (US)
Assigned to REGENTS OF THE UNIVERSITY OF MINNESOTA, Minneapolis, MN (US)
Filed by Regents of the University of Minnesota, Minneapolis, MN (US)
Filed on Jun. 10, 2020, as Appl. No. 16/897,977.
Claims priority of provisional application 62/859,864, filed on Jun. 11, 2019.
Prior Publication US 2020/0393445 A1, Dec. 17, 2020
Int. Cl. G01N 21/64 (2006.01); G01N 33/50 (2006.01)
CPC G01N 33/502 (2013.01) [G01N 21/6428 (2013.01); G01N 2021/6439 (2013.01); G01N 2333/4712 (2013.01)] 20 Claims
OG exemplary drawing
 
1. A method for identifying a compound that alters fluorescence resonance energy transfer (FRET) of a protein comprising:
providing a genetically engineered cell comprising a first protein and a second protein,
wherein the first protein comprises a first heterologous domain that comprises a first probe, and the first protein comprises either a CH domain-containing actin-binding domain or a tandem-CH, actin-binding domain,
wherein the second protein comprises a second heterologous domain that comprises a second probe;
wherein each protein binds to an actin filament in the cell in a spatial proximity to result in detectable FRET between the first and second probes;
contacting the cell with a test compound to form a mixture; and
measuring the fluorescence lifetime of the first probe, the second probe, or the combination thereof.