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Generation of Stable Expression Mammalian Cell Lines Using Lentivirus
Neha Tandon1, Kaushik N Thakkar2, Edward L LaGory2
1Department of Biology and Biochemistry, University of Houston, Houston, USA.
Bio-Protocol
|December 4, 2018
Summary
Third-generation lentiviruses provide a safe and robust method for creating stable mammalian cell lines for gene expression studies. This technique enables both gene down-regulation using short hairpin RNA (shRNA) and gene up-regulation via open reading frames (ORF).
Area of Science:
- Molecular Biology
- Cell Biology
- Gene Therapy
Background:
- Lentiviruses are widely employed for generating stable expression in mammalian cell lines.
- Applications include gene down-regulation (shRNA) and gene up-regulation (ORF).
Purpose of the Study:
- To highlight the utility and advantages of using third-generation lentiviruses for stable cell line generation.
- To emphasize the safety and efficiency of this molecular biology technique.
Main Methods:
- Utilizing third-generation lentivirus vectors for stable gene integration.
- Employing shRNA for gene knockdown and ORF for gene overexpression.
Main Results:
- The generation of stable mammalian cell lines using third-generation lentivirus is a robust technique.
- Stable expression is typically achieved within 1-2 weeks for most mammalian cell lines.
Conclusions:
- Third-generation lentiviruses offer a safe and replication-incompetent system for stable cell line development.
- This method provides a reliable approach for both gene silencing and overexpression studies in mammalian cells.
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