Modelling Intratumoral Treatment with miRNA Inhibitors: A Method to Analyze the Effect of miRNA Inhibitors on Tumor Growth in Murine Model
Published: April 30, 2023
Abstract
Source: Ramirez-Moya, J. et al. In Vivo Inhibition of MicroRNA to Decrease Tumor Growth in Mice. J. Vis. Exp. (2019)
This video demonstrates the in vivo delivery of miRNA inhibitors as therapeutic agents against tumor growth in orthotopic mouse models. The intratumoral administration of synthetic miRNA inhibitors or antagomiRs allows them to bind and inhibit endogenous miRNAs, thereby causing mRNA upregulation and suppression of tumor growth.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Flank Inoculation of Cells and Intratumoral antagomiR Treatment Cell preparation Engineer a Cal62 human thyroid cancer cell line (KRASG12R and p53A161D mutations) to overexpress a transgenic construct that constitutively expresses GFP and luciferase (CMV-Firefly Luc-IRES-eGFP). Select the transgenic cells by…
Divulgaciones
The authors have nothing to disclose.
Materials
| Basement Membrane Matrix: Matrigel Basement Membrane Matrix High Concentration | Corning | #354248 | |
| AntagomiR: mirVana miRNA inhibitor | Thermo Fisher | 4464088 | In vivo ready |
| In vivo delivery reagent: Invivofectamine 3.0 Reagent | Thermo Fisher | IVF 3005 | |
| Negative control: mirVana miRNA Inhibitor, Negative Control #1 | Thermo Fisher | 4464077 | In vivo ready |
| XenoLight D-Luciferin – K+ Salt Bioluminescent Substrate | PerkinElmer | 122799 | Diluted in PBS |
| In vivo imaging software: IVISLumina II Imaging System | Caliper Life Sciences |
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Modelling Intratumoral Treatment with miRNA Inhibitors: A Method to Analyze the Effect of miRNA Inhibitors on Tumor Growth in Murine Model. J. Vis. Exp. (Pending Publication), e20479, doi: (2023).