Using crispr/cas9 to knock out gm-csf in car-t cells

Rosalie M. Sterner, Michelle J. Cox, Reona Sakemura, Saad S. Kenderian

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

Chimeric antigen receptor T (CAR-T) cell therapy is a cutting edge and potentially revolutionary new treatment option for cancer. However, there are significant limitations to its widespread use in the treatment of cancer. These limitations include the development of unique toxicities such as cytokine release syndrome (CRS) and neurotoxicity (NT) and limited expansion, effector functions, and anti-tumor activity in solid tumors. One strategy to enhance CAR-T efficacy and/or control toxicities of CAR-T cells is to edit the genome of the CAR-T cells themselves during CAR-T cell manufacturing. Here, we describe the use of CRISPR/Cas9 gene editing in CAR-T cells via transduction with a lentiviral construct containing a guide RNA to granulocyte macrophage colony-stimulating factor (GM-CSF) and Cas9. As an example, we describe CRISPR/Cas9 mediated knockout of GM-CSF. We have shown that these GM-CSFk/o CAR-T cells effectively produce less GM-CSF while maintaining critical T cell function and result in enhanced anti-tumor activity in vivo compared to wild type CAR-T cells.

Original languageEnglish (US)
Article numbere59629
JournalJournal of Visualized Experiments
Volume2019
Issue number149
DOIs
StatePublished - Jul 2019

Keywords

  • CAR-T cell
  • CRISPR/Cas9
  • Cancer Research
  • Chimeric antigen receptor T cell
  • GM-CSF
  • Gene editing
  • Issue 149
  • Knockout

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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