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A Non-integrating Lentiviral Approach Overcomes Cas9-Induced Immune Rejection to Establish an Immunocompetent Metastatic Renal Cancer Model.

  • Author(s): Hu, Junhui
  • Schokrpur, Shiruyeh
  • Archang, Maani
  • Hermann, Kip
  • Sharrow, Allison C
  • Khanna, Prateek
  • Novak, Jesse
  • Signoretti, Sabina
  • Bhatt, Rupal S
  • Knudsen, Beatrice S
  • Xu, Hua
  • Wu, Lily
  • et al.
Abstract

The CRISPR-based technology has revolutionized genome editing in recent years. This technique allows for gene knockout and evaluation of function in cell lines in a manner that is far easier and more accessible than anything previously available. Unfortunately, the ability to extend these studies to in vivo syngeneic murine cell line implantation is limited by an immune response against cells transduced to stably express Cas9. In this study, we demonstrate that a non-integrating lentiviral vector approach can overcome this immune rejection and allow for the growth of transduced cells in an immunocompetent host. This technique enables the establishment of a von Hippel-Lindau (VHL) gene knockout RENCA cell line in BALB/c mice, generating an improved model of immunocompetent, metastatic renal cell carcinoma (RCC).

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