Related Experiment Video
Updated: Dec 30, 2025
In Vitro Establishment of a Genetically Engineered Murine Head and Neck Cancer Cell Line using an Adeno-Associated Virus-Cas9 System
Published on: January 9, 2020
In Vitro Establishment of a Genetically Engineered Murine Head and Neck Cancer Cell Line using an Adeno-Associated
Manu Prasad1, Sankar Jagadeeshan1, Maurizio Scaltriti2
1The Shraga Segal Department of Microbiology, Immunology and Genetics, Ben-Gurion University of the Negev; Faculty of Health Sciences, Ben-Gurion University of the Negev.
Abstract:
The use of primary normal epithelial cells makes it possible to reproducibly induce genomic alterations required for cellular transformation by introducing specific mutations in oncogenes and tumor suppressor genes, using clustered regulatory interspaced short palindromic repeat (CRISPR)-based genome editing technology in mice. This technology allows us to accurately mimic the genetic changes that occur in human cancers using mice. By genetically transforming murine primary cells, we can better study cancer development, progression, treatment, and diagnosis. In this study, we used Cre-inducible Cas9 mouse tongue epithelial cells to enable genome editing using adeno-associated virus (AAV) in vitro. Specifically, by altering KRAS, p53, and APC in normal tongue epithelial cells, we generated a murine head and neck cancer (HNC) cell line in vitro,which is tumorigenic in syngeneic mice. The method presented here describes in detail how to generate HNC cell lines with specific genomic alterations and explains their suitability for predicting tumor progression in syngeneic mice. We envision that this promising method will be informative and useful to study tumor biology and therapy of HNC.
Insights
Researchers created a new mouse model for head and neck cancer (HNC) by precisely editing genes in normal tongue cells. This model accurately mimics human cancer genetics, aiding in the study of tumor development and treatment.
Area of Science:
- Oncology
- Genetics
- Molecular Biology
Background:
- Cancer research often requires accurate models to study disease progression and test therapies.
- Genetic alterations in oncogenes and tumor suppressor genes are fundamental to cancer development.
- Existing models may not fully recapitulate the specific genetic changes observed in human cancers.
Purpose of the Study:
- To develop a reproducible method for generating genetically engineered head and neck cancer (HNC) cell lines in vitro.
- To utilize clustered regularly interspaced short palindromic repeats (CRISPR)-based genome editing for precise genetic modification of primary cells.
- To establish a murine model for studying HNC development, progression, and potential therapeutic strategies.
Main Methods:
- Primary mouse tongue epithelial cells were genetically modified using CRISPR technology.
- Specific mutations were introduced into key genes such as KRAS, p53, and APC.
- Genome editing was performed in vitro using Cre-inducible Cas9 and adeno-associated virus (AAV) delivery.
- The generated cell line was tested for tumorigenicity in syngeneic mice.
Main Results:
- A tumorigenic murine head and neck cancer (HNC) cell line was successfully generated in vitro.
- The cell line harbors specific, engineered genomic alterations mimicking human cancer genetics.
- The generated HNC cells demonstrated tumor formation when implanted in syngeneic mice.
- The study details the methodology for creating these specific cell lines.
Conclusions:
- The described method provides a robust approach for generating HNC cell lines with defined genetic alterations.
- This in vitro model system is suitable for studying tumor biology and predicting tumor progression.
- The model holds promise for advancing the understanding and treatment of head and neck cancers.
