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

Molecular Therapy. Methods & Clinical Development
Junhui HuLily Wu

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).

Citations

Dec 1, 2019·Nature Communications·Krzysztof ChylinskiUlrich Elling
Sep 15, 2020·FASEB BioAdvances·Natasha L FriendAndrew C W Zannettino
Sep 5, 2020·The Journal of Biological Chemistry·Abigail AllenPartha Roy
Feb 10, 2021·Signal Transduction and Targeted Therapy·Jote T BulchaGuangping Gao
Feb 25, 2020·Translational Oncology·Paweł SobczukAnna M Czarnecka
Mar 11, 2021·Immunity·Fei Yi, Christopher A Klebanoff
Apr 5, 2021·Trends in Immunology·Frank A Buquicchio, Ansuman T Satpathy

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Methods Mentioned

BETA
transfection
ELISA
Knockout
light microscopy
gene knockout
genetic modification
transgenic

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