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JoVE Journal

Bioengineering

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An Integrated System to Remotely Trigger Intracellular Signal Transduction by Upconversion Nanoparticle-mediated Kinase Photoactivation

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11:20 min

August 30th, 2017

August 30th, 2017

7,317 Views

0:05

Title

1:33

Fluorimeter Setup for Upconversion Spectrum Measurement and Microscope Setup

3:29

Characterization of Caged-PKA and Caged PKA/UCNP Constructs and Photolysis Setup

4:55

Cell Sample Preparation and Microinjection of Caged-PKA/UCNP Complexes

7:35

Photoactivation of Caged-PKA/UCNP Using NIR Light in Living Cells Experiment

8:33

Results: Visualize PKA Signal Transduction Activation via NIR Photoactivation

9:40

Conclusion

Transcript

The goal of this experiment is to demonstrate the feasibility to photoactivate enzyme using near infrared irradiation. Caged-protein kinase A is immobilized on this surface of upconversion nanoparticles micro-injected into cells and kinase induced

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In this protocol, caged protein kinase A (PKA), a cellular signal transduction bioeffector, was immobilized on a nanoparticle surface, microinjected into the cytosol, and activated by the upconverted UV light from near-infrared (NIR) irradiation, inducing downstream stress fiber disintegration in the cytosol.

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