Intracardiac Delivery of Viral Vectors: A Technique to Inject Viral Vectors Intracardially to Study Pancreatic Metastasis in Murine Model
Published: April 30, 2023
Abstract
Source: Zhang, G., et al. Identification and Characterization of Metastatic Factors by Gene Transfer into the Novel RIP-Tag; RIP-tva Murine Model. J. Vis. Exp. (2017)
This video describes the protocol for intracardiac injection of retroviruses into the transgenic mouse to ensure gene transfer into proliferating pancreatic β premalignant lesions. This method can also help in assessing the functions of candidate genes in tumor metastasis.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the jove veterinary review board. 1. Intracardiac injection: Hemizygous RIP-Tag mice, not homozygous mice, are used in this study. Hemizygous RIP-Tag mice develop pancreatic neuroendocrine tumors around 10~12 weeks of age, while homozygous mice have a shorter tumor latency. Because cell proliferation is requi…
Representative Results
Figure 1: Placement of intracardiac injection. Anatomical landmarks are shown with horizontal dashed lines on the sternum (outlined in white). On the mouse's skin, the sternal notch and xiphoid process serve as landmarks, and the needle is inserted 1 mm away from the mid-sternum and slightly left (anatomical) of the sternum. <a href="https://www.jove.com/files/ftp_upload/203…
Divulgazioni
The authors have nothing to disclose.
Materials
| Insulin Syringes | BD | 329461 | |
| RCASBP-Y DV plasmid | Addgene | 11478 | |
| RCAS-RNAi plasmid | Addgene | 15182 | |
| DMEM | Corning | 10-013-CV | |
| Penicillin-Streptomycin solution, 100x | Corning | 30-002-CI | |
| Superfect | Qiagen | 301305 | |
| 0.45 mm Nalgene Syringe Filters with PES Membrane | Thermo Scientific | 194-2545 | |
| Synaptophysin | Vector Laboratories | VP-S284 | |
| VECTASTAIN Elite ABC HRP Kit (Peroxidase, Rabbit IgG) | Vector Laboratories | PK-6101 | |
| AmpliTaq DNA Polymerase with Buffer II | Life Technologies | N8080153 | |
| PBS-/-, 1x | Corning | 21-040-CV | |
| Superfect | Qiagen | 301305 | |
| Synaptophysin | Vector Laboratories | VP-S284 | |
| L-glutamine, 100x | Corning | 25-005-CI | |
| Fetal bovine serum | Atlanta Biologicals | 25-005-CI |
Citazione di questo articolo
Intracardiac Delivery of Viral Vectors: A Technique to Inject Viral Vectors Intracardially to Study Pancreatic Metastasis in Murine Model. J. Vis. Exp. (Pending Publication), e20317, doi: (2023).