TY - JOUR
T1 - Rapid biosensor development using plant hormone receptors as reprogrammable scaffolds
AU - Beltrán, Jesús
AU - Steiner, Paul J.
AU - Bedewitz, Matthew
AU - Wei, Shuang
AU - Peterson, Francis C.
AU - Li, Zongbo
AU - Hughes, Brigid E.
AU - Hartley, Zachary
AU - Robertson, Nicholas R.
AU - Medina-Cucurella, Angélica V.
AU - Baumer, Zachary T.
AU - Leonard, Alison C.
AU - Park, Sang Youl
AU - Volkman, Brian F.
AU - Nusinow, Dmitri A.
AU - Zhong, Wenwan
AU - Wheeldon, Ian
AU - Cutler, Sean R.
AU - Whitehead, Timothy A.
N1 - Funding Information:
This work was supported by the Defense Advanced Research Projects Agency Advanced Plant Technologies (grant HR001118C0137 to D.A.N., T.A.W., S.R.C. and I.W.), the National Science Foundation (grant NSF- 2128287 to T.A.W., grant NSF-2128016 to S.R.C. and I.W. and grant NSF-2128246 to F.C.P.), the National Science Foundation Graduate Research Fellowship Program (A.C.L. and Z.T.B.) and the National Institutes of Health National Institute on Drug Abuse (grant 1R21DA053496-01 to S.R.C and W.Z.). The views, opinions, and/or findings expressed are those of the authors and should not be interpreted as representing the official views or policies of the Department of Defense or the U.S. Government. Approved for Public Release, Distribution Unlimited. Z.H. and N.R.R. were supported by the National Science Foundation Plants3D NRT project (grant NSF-1922642). Additional support for F.P. was provided by the Medical College of Wisconsin.
Funding Information:
This work was supported by the Defense Advanced Research Projects Agency Advanced Plant Technologies (grant HR001118C0137 to D.A.N., T.A.W., S.R.C. and I.W.), the National Science Foundation (grant NSF- 2128287 to T.A.W., grant NSF-2128016 to S.R.C. and I.W. and grant NSF-2128246 to F.C.P.), the National Science Foundation Graduate Research Fellowship Program (A.C.L. and Z.T.B.) and the National Institutes of Health National Institute on Drug Abuse (grant 1R21DA053496-01 to S.R.C and W.Z.). The views, opinions, and/or findings expressed are those of the authors and should not be interpreted as representing the official views or policies of the Department of Defense or the U.S. Government. Approved for Public Release, Distribution Unlimited. Z.H. and N.R.R. were supported by the National Science Foundation Plants3D NRT project (grant NSF-1922642). Additional support for F.P. was provided by the Medical College of Wisconsin.
Publisher Copyright:
© 2022, The Author(s).
PY - 2022/12
Y1 - 2022/12
N2 - A general method to generate biosensors for user-defined molecules could provide detection tools for a wide range of biological applications. Here, we describe an approach for the rapid engineering of biosensors using PYR1 (Pyrabactin Resistance 1), a plant abscisic acid (ABA) receptor with a malleable ligand-binding pocket and a requirement for ligand-induced heterodimerization, which facilitates the construction of sense–response functions. We applied this platform to evolve 21 sensors with nanomolar to micromolar sensitivities for a range of small molecules, including structurally diverse natural and synthetic cannabinoids and several organophosphates. X-ray crystallography analysis revealed the mechanistic basis for new ligand recognition by an evolved cannabinoid receptor. We demonstrate that PYR1-derived receptors are readily ported to various ligand-responsive outputs, including enzyme-linked immunosorbent assay (ELISA)-like assays, luminescence by protein-fragment complementation and transcriptional circuits, all with picomolar to nanomolar sensitivity. PYR1 provides a scaffold for rapidly evolving new biosensors for diverse sense–response applications.
AB - A general method to generate biosensors for user-defined molecules could provide detection tools for a wide range of biological applications. Here, we describe an approach for the rapid engineering of biosensors using PYR1 (Pyrabactin Resistance 1), a plant abscisic acid (ABA) receptor with a malleable ligand-binding pocket and a requirement for ligand-induced heterodimerization, which facilitates the construction of sense–response functions. We applied this platform to evolve 21 sensors with nanomolar to micromolar sensitivities for a range of small molecules, including structurally diverse natural and synthetic cannabinoids and several organophosphates. X-ray crystallography analysis revealed the mechanistic basis for new ligand recognition by an evolved cannabinoid receptor. We demonstrate that PYR1-derived receptors are readily ported to various ligand-responsive outputs, including enzyme-linked immunosorbent assay (ELISA)-like assays, luminescence by protein-fragment complementation and transcriptional circuits, all with picomolar to nanomolar sensitivity. PYR1 provides a scaffold for rapidly evolving new biosensors for diverse sense–response applications.
UR - http://www.scopus.com/inward/record.url?scp=85132543880&partnerID=8YFLogxK
U2 - 10.1038/s41587-022-01364-5
DO - 10.1038/s41587-022-01364-5
M3 - Article
C2 - 35726092
AN - SCOPUS:85132543880
SN - 1087-0156
VL - 40
SP - 1855
EP - 1861
JO - Nature Biotechnology
JF - Nature Biotechnology
IS - 12
ER -