In vitro establishment of a genetically engineered murine head and neck cancer cell line using an adeno-associated Virus-Cas9 system

Manu Prasad, Sankar Jagadeeshan, Maurizio Scaltriti, Irit Allon, Moshe Elkabets

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

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.

Original languageEnglish
Article numbere60410
JournalJournal of Visualized Experiments
Volume2020
Issue number155
DOIs
StatePublished - 1 Jan 2019

Keywords

  • Adeno-associated virus vector
  • CRISPR
  • Cancer Research
  • Genomic editing
  • Head and neck cancer
  • In vitro cell transformation
  • Issue 155
  • Murine cancer models
  • Tumor cell line

ASJC Scopus subject areas

  • General Neuroscience
  • General Chemical Engineering
  • General Biochemistry, Genetics and Molecular Biology
  • General Immunology and Microbiology

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