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Reliably Engineering and Controlling Stable Optogenetic Gene Circuits in Mammalian Cells
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Chapitres
- 00:04Introduction
- 00:43Gene Circuit Design
- 01:39Stable Cell Line Engineering
- 02:27Light Plate Apparatus Induction Assays
- 04:08Fluorescence Microscopy of Light-Induced Engineered Cells
- 04:32Flow Cytometry of Light-Induced Engineered Cells
- 05:14Immunofluorescence of Gene Circuit Components
- 07:16Results: Gene Expression Analysis in Engineered Optogenetic Cell Lines
- 08:49Conclusion
Reliably controlling light-responsive mammalian cells requires the standardization of optogenetic methods. Toward this goal, this study outlines a pipeline of gene circuit construction, cell engineering, optogenetic equipment operation, and verification assays to standardize the study of light-induced gene expression using a negative-feedback optogenetic gene circuit as a case study.
