In Vivo CRISPR Screening Case Study I Pharmaron CRO

Poster on in vivo CRISPR screening for therapeutic target identification using a xenograft model, with charts, graphs, and downloadable PDF option.

Poster Authors:

Ce Zhong, Lu Bai, Tongtong Wang, Yiying Shao, Lingyun Zhang, Lu Zhang, Jingqi Huang

¹in vivo Pharmacology, Pharmaron Beijing Co., Ltd. (China)

in vivo CRISPR Screening Identifies Disease Drivers in the Tumor Environment

Conventional target identification approaches are often performed in vitro and may fail to fully capture the physiological complexities of a tumor. Through in vivo CRISPR screening, Pharmaron scientists perturb thousands of genes simultaneously in tumors growing in animals to reveal drivers of tumor growth and drug resistance.

Which information can be gained with in vivo CRISPR screening?

Unbiased evaluation of thousands of genes in a single study
Evaluation of gene function in the physiological tumor environment
Identification of resistance mechanisms to drug treatment
Discovery of biomarkers associated with drug resistance
Identification of targets for combination treatment

Case Study of in vivo CRISPR Screening to Identify Genes that Influence the Response to Trametinib Treatment

The poster shows how in vivo CRISPR screening is used to identify genes whose knockdown increases or decreases tumor cell survival during treatment of the MEK1/2 inhibitor trametinib.

Capabilities

Knowledge Center

in vivo Pharmacology

Oncology Animal Models

Connect with a Scientific Expert

Complete the form to access resource

Name(Required)

First Last

Company Name(Required)

Job Title(Required)

Country(Required)

State(Required)

Message

Email(Required)

This field is hidden when viewing the form

Lead Source(Required)

By completing and submitting this form, you acknowledge that you have read and understood our Privacy Policy, and you consent to the collection and processing of your personal data by Pharmaron for the purpose of responding to your inquiry. You also agree to be contacted by Pharmaron regarding relevant services and updates. You may withdraw your consent at any time.

Study Objectives:

Identify genes whose loss causes tumor cells to disappear during Trametinib treatment, indicating targets for combination treatment.
Identify genes whose loss causes tumor cells to better survive during Trametinib treatment, contributing to drug resistance.

Study Design: in vivo CRISPR Screening under Drug Pressure

A549 human lung tumor cells were transfected with 3,188 single guide RNAs (sgRNA) targeting tumor suppressor genes, with one sgRNA per cell. The study design comprised three study groups:

Tumors implanted in mice and subsequently treated with either vehicle or Trametinib
Tumor cells grown in vitro cell culture as engineered controls

After three weeks, the tissues were amplicon sequenced to identify residual sgRNAs, which represent genes that provided a survival advantage.

Study Results

The screen identified multiple genes when comparing in vitro-grown cells with in vivo-grown tumors (see table A), as well as when comparing vehicle versus- and trametinib-treated tumors (see table B). The screen identified genes whose knockout either promoted or impaired tumor growth under the pressure of the in vivo physiologic environment (A) or treatment pressure (B).

Explore the full study design, gene enrichment analysis, and identified targets by downloading the poster.