A CRISPR/Cas9-Based Assay for High-Throughput Studies of Cancer-Induced Innervation
Abstract
:Simple Summary
Abstract
1. Introduction
2. Results
2.1. Cancer-Induced Neuronal Precursor Differentiation in Coculture In Vitro
2.2. Endogenous Labeling of β3-Tubulin Microtubule Protein in Neuronal Precursors
2.3. Characterization of the TUBB3-GFP Knock-in Approach
2.4. Validation of the Reporter Approach for High-Throughput Study of Cancer-Induced Neuronal Differentiation in Direct Coculture In Vitro
2.5. Breast Cancer Plasticity Controls Neuronal Precursors Differentiation
3. Discussion
4. Methods
4.1. Cell Culture and Reagents
4.2. Generation of PC12-GFP and PC12 mCherry
4.3. Generation of PC12-tubb3-GFP/ Neuro2A-tubb3-GFP Cell Lines
4.4. Flow Cytometry
4.5. Western Blot Analysis
4.6. Direct Counting Cell Proliferation Assay
4.7. WST1 Metabolic Activity Assay
4.8. Migration Assay
4.9. Fluorimeter and Microscopy Analysis
4.10. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Galappaththi, S.L.; Katz, B.; Howze, P.H., IV; Hoover, G.; Grelet, S. A CRISPR/Cas9-Based Assay for High-Throughput Studies of Cancer-Induced Innervation. Cancers 2023, 15, 2026. https://doi.org/10.3390/cancers15072026
Galappaththi SL, Katz B, Howze PH IV, Hoover G, Grelet S. A CRISPR/Cas9-Based Assay for High-Throughput Studies of Cancer-Induced Innervation. Cancers. 2023; 15(7):2026. https://doi.org/10.3390/cancers15072026
Chicago/Turabian StyleGalappaththi, Sapthala Loku, Brenna Katz, Patrick H. Howze, IV, Gregory Hoover, and Simon Grelet. 2023. "A CRISPR/Cas9-Based Assay for High-Throughput Studies of Cancer-Induced Innervation" Cancers 15, no. 7: 2026. https://doi.org/10.3390/cancers15072026