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Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
Published on: June 16, 2017
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Avian Retrovirus-Mediated Tumor-Specific Gene Knockout
Wei Wang1, Bingning Dong1, Feng Yang1
1Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas.
Current Protocols in Molecular Biology
|January 17, 2018
Summary
We developed a novel gene delivery system, RCAS-Cre-IRES-Oncogene (RCI-Oncogene), for in vivo tumor gene knockout. This system demonstrated FGFR1
Area of Science:
- Oncology
- Gene Therapy
- Cancer Research
Background:
- The replication-competent avian sarcoma leukosis virus long-terminal repeat with splice acceptor-tumor virus A (RCAS-TVA) system is established for cancer modeling.
- Spontaneous tumor gene manipulation in vivo requires efficient and versatile gene delivery methods.
Purpose of the Study:
- To introduce a novel gene delivery system, RCAS-Cre-IRES-Oncogene (RCI-Oncogene), for precise gene manipulation in spontaneous tumors.
- To establish a tumor gene knockout (TuKO) protocol using the RCI-Oncogene system.
- To investigate the role of FGFR1 in mammary tumor metastasis.
Main Methods:
- Development of the RCI-Oncogene system for efficient in vivo gene delivery.
- Application of the RCI-Oncogene system for tumor gene knockout (TuKO).
- Utilizing mutant mouse hosts for targeted gene modification in tumors.
Main Results:
- The RCI-Oncogene system enables efficient manipulation of gene expression in spontaneous tumors.
- The TuKO approach using RCI-Oncogene demonstrated a critical role for FGFR1 in mammary tumor metastasis.
Conclusions:
- The RCI-Oncogene system is a versatile tool for tumor gene modification in vivo.
- The TuKO approach provides a robust method for studying gene function in spontaneous tumors.
- This technology can be adapted for various research questions in appropriate mouse models.
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