Virus Delivery of CRISPR Guides to the Murine Prostate for Gene Alteration

Maria Riedel; Martin F. Berthelsen; Latifa Bakiri; Erwin F. Wagner; Martin K. Thomsen

doi:10.3791/57525

Virus Delivery of CRISPR Guides to the Murine Prostate for Gene Alteration

JoVE Journal
Cancer Research

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JoVE Journal Cancer Research

Virus Delivery of CRISPR Guides to the Murine Prostate for Gene Alteration

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06:21 min

April 27, 2018

DOI: 10.3791/57525-v

Maria Riedel¹, Martin F. Berthelsen¹, Latifa Bakiri², Erwin F. Wagner², Martin K. Thomsen¹

¹Department of Clinical Medicine,Aarhus University, ²GDD, Cancer Cell Biology Program,National Cancer Research Center (CNIO)

Overview

This protocol describes a method for virus delivery to the murine prostate, utilizing CRISPR/Cas9 technology for gene editing. This technique enables orthotopic alteration of gene expression and introduces a novel mouse model for studying prostate cancer.

Key Study Components

Area of Science

Neuroscience
Gene Editing
Prostate Cancer Research

Background

The surgical procedure aims to alter gene expression in the murine prostate.
Viral delivery of CRISPR/Cas9 guides is employed for prostate cancer induction.
This method addresses new questions in prostate cancer biology.
It allows for the analysis and validation of new genes in an in vivo setting.

Purpose of Study

To enable rapid gene editing of multiple genes.
To target a subset of prostatic cells, mimicking human scenarios.
To improve understanding of gene functions in prostate cancer.

Methods Used

Preparation of anesthetic for surgical procedures.
Viral delivery techniques for gene editing.
CRISPR/Cas9 technology for precise gene alteration.
Use of a novel mouse model for in vivo studies.

Main Results

Successful alteration of gene expression in murine prostate.
Demonstrated potential for rapid gene editing.
Validated the effectiveness of the new mouse model.
Provided insights into prostate cancer biology.

Conclusions

The method offers a promising approach for prostate cancer research.
It enhances the ability to study gene functions in vivo.
Future studies can build on this technique to explore new therapeutic avenues.

Frequently Asked Questions

What is the main goal of this protocol?

The main goal is to alter gene expression in the murine prostate using viral delivery methods.

What advantages does this method offer?

It allows rapid gene editing of multiple genes by targeting specific prostatic cells.

What challenges might a beginner face?

Beginners may struggle with the organ size and the precision required for successful surgery.

How is the anesthetic prepared for the procedure?

Mix specific volumes of medetomidine, midazolam, butorphanol, and sterile saline.

What technology is primarily used in this study?

CRISPR/Cas9 technology is primarily used for gene editing.

What is the significance of the new mouse model?

It provides a novel platform for studying prostate cancer in vivo.

This protocol describes a newly established method for virus delivery to the murine prostate. Using either CRISPR/Cas9 technology, gene overexpression, or Cre recombinase delivery, the technique allows orthotopic alteration of gene expression and implements a novel mouse model for prostate cancer.

The overall goal of this surgical procedure is to alter gene expression in the murine prostate by viral delivery of CRISPR Cas9 guides for prostate cancer induction. This method can help addressing new questions in prostate cancer biology regards to analyzing and validating new genes in an in vivo setting. The main advantages of this technique is it allow rapid gene editing of multiple genes by only targeting a subset of the prostatic cells as seen in the human scenario.

Someone who is new to the procedure may struggle at first because of organ size, and the precision that is needed for a successful surgical completion. To freshly prepare a total volume of 5 milliliters of anesthetic, mix 0.15 milliliters of one mg per milliliter medetomidine hydrochloride, 0.4 milliliters of 5 mg per milliliter of midazolam, and 0.25 milliliters of 10 mg per milliliter butorphenol. Then, add 4.2 milliliters of sterile saline water.

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