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)
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.