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Defining Gene Functions in Tumorigenesis by Ex vivo Ablation of Floxed Alleles in Malignant Peripheral Nerve Sheath Tumor Cells
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Cancer Research
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JoVE Journal Cancer Research
Defining Gene Functions in Tumorigenesis by Ex vivo Ablation of Floxed Alleles in Malignant Peripheral Nerve Sheath Tumor Cells

Defining Gene Functions in Tumorigenesis by Ex vivo Ablation of Floxed Alleles in Malignant Peripheral Nerve Sheath Tumor Cells

DOI:
10.3791/62740-v

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09:37 min

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August 25, 2021

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, ,
1Department of Pathology and Laboratory Medicine,Medical University of South Carolina, 2Department of Pediatrics and Neurology,Perelman School of Medicine of the University of Pennsylvania, 3Division of Child Neurology,Children’s Hospital of Philadelphia

Chapters

  • 00:05Introduction
  • 00:52Tumor Dissection and Staining
  • 01:46Ex Vivo Ablation of Floxed Erbb4 Alleles in MPNST Cells
  • 03:52Proliferation Assays in MPNST Cells with Ablated Erbb4 Alleles
  • 04:23RNA-Seq Analyses and Gene Identification with Altered Expression by Erbb4 Loss
  • 06:04Orthotopic Allograft of MPNST Cells with Ablated Erbb4 Alleles and Erbb4 Ablation Effects Analysis
  • 07:10Results: Effect of Adenovirus Transduction on MPNST Cells and Allografts
  • 09:09Conclusion

Summary

Automatic Translation

Automatic Translation

Here, we present a protocol for performing gene knockouts that are embryonic lethal in vivo in genetically engineered mouse model-derived tumors and then assessing the effect that the knockout has on tumor growth, proliferation, survival, migration, invasion, and the transcriptome in vitro and in vivo.

Tags

Keywords: Gene FunctionTumorigenesisEx Vivo AblationFloxed AllelesMalignant Peripheral Nerve Sheath TumorGenetically Engineered Mouse ModelAdenoviral InfectionEarly Passage CultureImmunohistochemical StainingNeuregulin-1 BetaForskolinEGFP

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