Modeling Prostate Cancer in Genetically-engineered Mouse Models: A CRISPR/Cas9-mediated Localized Gene Editing Technique in Mouse Anterior Prostate Lobe Cells

Published: April 30, 2023

Abstract

Source: Riedel, M. et al. Virus Delivery of CRISPR Guides to the Murine Prostate for Gene Alteration. J. Vis. Exp. (2018)

This video describes a method for editing specific genes in prostate gland cells using an adenovirus-based delivery system. The approach allows for orthotopic and localized alteration of gene expression utilizing CRISPR technology to develop novel mouse models for prostate cancer.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Virus-delivery to the Murine Prostate Anesthetize the animal by intraperitoneal injection with a sterile 1 mL syringe and a 27G x ½" needle. Use an anesthesia dose of 0.01 mL/g body weight. If the procedure lasts longer than 30 min, maintain anesthesia by injecting 1/3 of the starting dose every 30 min. <…

Representative Results

Figure 1: Illustration of the procedure. The procedure is carried out in the Rosa26-LSL-Cas9-EGFP mouse generated by Platt et al. The anterior prostate (AP) is attached to the seminal vesicle (SV). Virus particles expressing guide RNA and a Cre protein are injected into the anterior prostate to alter gene expression.

Declarações

The authors have nothing to disclose.

Materials

B6J.129(B6N)-Gt(ROSA)26Sor^{tm1(CAG-cas9*,-}
^{EGFP)Fezh} /J mice
The Jackson Laboratory 26175
0,5 ml U-100 insulin syringe BD 324825 for virus injection
1 ml syringe BD 300013 for anesthesia injection
30 G 1/2'' needle BD 304000 for anesthesia injection
6-0 Polysorb Suture Medtronic GL889 to close the peritoneum
Disposable sterilized surgery drape multiple suppliers
Heating pad multiple suppliers
Povidone-Iodine Prep Pad Fisher Scientific 06-669-70
Trimming machine Aesculap GT415 to shave the abdomen
Dumont Forceps F.S.T 11252-00
Halsey Micro Needle Holder F.S.T 12500-12
Iris Scissors F.S.T 14094-11
Narrow Pattern Forceps F.S.T 11002-12
Ring Forceps F.S.T 11106-09
Wound Clip System Handle incl.
Clips
F.S.T 12030-01 to close the skin
Wound Clip System Remover F.S.T 12030-04
1x PBS Gibco 10010-023
Antisedan obtained from the animal facility
Butorphanol obtained from the animal facility
Medetomidinhydrochlorid obtained from the animal facility
Midazolam obtained from the animal facility
100% Ethanol Fisher Scientific 22-032-103
Eye ointment Takeda 7242
Virus (AAV, AV) multiple suppliers virus used in this protocol is an in-house production

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Citar este artigo
Modeling Prostate Cancer in Genetically-engineered Mouse Models: A CRISPR/Cas9-mediated Localized Gene Editing Technique in Mouse Anterior Prostate Lobe Cells. J. Vis. Exp. (Pending Publication), e20407, doi: (2023).

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