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Spatiotemporal precise optical manipulation of intracellular molecular activities

Bin Dong, Shivam Mahapatra, Matthew G. Clark, Mark Carlsen, Karsten J. Mohn, Seohee Ma, Kent Brasseale, Grace Crim, Chi Zhang
doi: https://doi.org/10.1101/2023.07.19.549752
Bin Dong
1Department of Chemistry, Purdue University, 560 Oval Dr., West Lafayette, IN 47907, USA
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Shivam Mahapatra
1Department of Chemistry, Purdue University, 560 Oval Dr., West Lafayette, IN 47907, USA
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Matthew G. Clark
1Department of Chemistry, Purdue University, 560 Oval Dr., West Lafayette, IN 47907, USA
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Mark Carlsen
1Department of Chemistry, Purdue University, 560 Oval Dr., West Lafayette, IN 47907, USA
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Karsten J. Mohn
1Department of Chemistry, Purdue University, 560 Oval Dr., West Lafayette, IN 47907, USA
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Seohee Ma
1Department of Chemistry, Purdue University, 560 Oval Dr., West Lafayette, IN 47907, USA
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Kent Brasseale
1Department of Chemistry, Purdue University, 560 Oval Dr., West Lafayette, IN 47907, USA
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Grace Crim
1Department of Chemistry, Purdue University, 560 Oval Dr., West Lafayette, IN 47907, USA
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Chi Zhang
1Department of Chemistry, Purdue University, 560 Oval Dr., West Lafayette, IN 47907, USA
2Purdue Center for Cancer Research, 201 S. University St., West Lafayette, IN 47907, USA
3Purdue Institute of Inflammation, Immunology, and Infectious Disease, 207 S. Martin Jischke Dr., West Lafayette, IN 47907, USA
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  • For correspondence: zhan2017@purdue.edu
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Abstract

Controlling chemical processes in live cells is a challenging task. The spatial heterogeneity of biochemical reactions in cells is often overlooked by conventional means of incubating cells with desired chemicals. A comprehensive understanding of spatially diverse biochemical processes requires precise control over molecular activities at the subcellular level. Herein, we develop a closed-loop optoelectronic control system that allows the manipulation of biomolecular activities in live cells at high spatiotemporal precision. Chemical-selective fluorescence signals are utilized to command lasers that trigger specific chemical reactions or control the activation of photoswitchable inhibitors at desired targets. We demonstrate the capability to selectively produce reactive oxygen species (ROS) solely at targeted organelles using blue light. Notably, the induction of ROS in the endoplasmic reticulum leads to a more pronounced disruption of tubulin polymerization and a reduction in green fluorescent protein signals, in comparison to that in lipid droplets. Moreover, when combined with a photoswitchable inhibitor, we selectively inhibit tubulin polymerization within subcellular compartments. This technology enables spatiotemporal control over chemical processes and drug activities, exclusively at desired targets, while minimizing undesired effects on non-targeted locations.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted July 20, 2023.
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Spatiotemporal precise optical manipulation of intracellular molecular activities
Bin Dong, Shivam Mahapatra, Matthew G. Clark, Mark Carlsen, Karsten J. Mohn, Seohee Ma, Kent Brasseale, Grace Crim, Chi Zhang
bioRxiv 2023.07.19.549752; doi: https://doi.org/10.1101/2023.07.19.549752
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Spatiotemporal precise optical manipulation of intracellular molecular activities
Bin Dong, Shivam Mahapatra, Matthew G. Clark, Mark Carlsen, Karsten J. Mohn, Seohee Ma, Kent Brasseale, Grace Crim, Chi Zhang
bioRxiv 2023.07.19.549752; doi: https://doi.org/10.1101/2023.07.19.549752

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