Rapid biosensor development using plant hormone receptors as reprogrammable scaffolds.
Jesús BeltránPaul J SteinerMatthew BedewitzShuang WeiFrancis C PetersonZongbo LiBrigid E HughesZachary HartleyNicholas R RobertsonAngélica V Medina-CucurellaZachary T BaumerAlison C LeonardSang-Youl ParkBrian F VolkmanDmitri A NusinowWenwan ZhongIan WheeldonSean R CutlerTimothy A WhiteheadPublished in: Nature biotechnology (2022)
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.
Keyphrases
- label free
- loop mediated isothermal amplification
- high throughput
- transcription factor
- sensitive detection
- quantum dots
- tissue engineering
- high resolution
- binding protein
- gold nanoparticles
- gene expression
- high glucose
- diabetic rats
- arabidopsis thaliana
- drug delivery
- oxidative stress
- protein protein
- amino acid
- heat stress
- heat shock
- magnetic resonance
- real time pcr
- dual energy
- heat shock protein