Lentivirus Transfection
BioInnovatise Viral Vector Team
Updated July 15, 2024
Lentivirus transfection is the process used to produce lentiviral particles during a process known as lentivirus packaging. But what exactly is lentivirus transfection? How does it differ with lentivirus transduction? Are there certain considerations that need to be made in order to increase your lentivirus transfection efficiency in your packaging protocol?
Our viral vector team has answered some of the frequent lentivirus transfection questions you may have when requesting lentivirus packaging services.
What's The Difference Between Lentivirus Transfection And Transduction?
Lentivirus Transfection:
- Transfection is a process used to introduce nucleic acids (DNA or RNA) into eukaryotic cells.
- Method: In the context of lentiviruses, transfection usually refers to the introduction of lentiviral vectors (i.e. packaging plasmids) into producer cells to produce lentiviral particles. There are many producer cell lines to choose from however HEK293T cell line is ideal for achieving a high transfection rate.
- The primary goal of transfection in this context is to produce lentiviral particles that can then be used for transduction.
Lentivirus Transduction:
- Transduction is the process by which viral particles transfer genetic material into target cells.
- Method: For lentiviruses, transduction involves the introduction of the viral particles (produced from the transfection of producer cells) into the target cells. The viral particles fuse with the target cell membrane, allowing the viral RNA to enter and integrate into the host cell genome.
- The primary goal of transduction is to achieve stable and efficient gene delivery and expression in the target cells.
The above diagram illustrates the lentivirus packaging process at BioInnovatise, where our team uses HEK293T cells for lentivirus packaging.
How Can You Increase Lentivirus Transfection Efficiency
Optimization of Transfection Reagents and Conditions:
- Transfection Reagent Choice:
- Use high-quality, efficient transfection reagents specifically designed for lentiviral vector production, such as Lipofectamine 2000 or 3000, PEI (polyethylenimine), or calcium phosphate. Lentivirus packaging mixes may include transfection reagents.
- DNA Quality:
- Use high-purity plasmid DNA to avoid contaminants that can inhibit transfection. Our team can increase your plasmid DNA’s quality with our molecular cloning services.
- Ensure your plasmid DNA has a low nucleic acid ratio. Learn about endotoxin free plasmid prep, as endotoxins can significantly reduce transfection efficiency.
- DNA Quantity and Ratio:
- Too much or too little DNA can negatively impact transfection efficiency.
- Use the optimal ratio of lentiviral plasmids i.e. transfer plasmid, packaging plasmid, and envelope plasmid. Common ratios are 4:3:1 or 3:2:1 depending on your lentivirus packaging generation.
Transfection is a pivotal step in the lentivirus packaging protocol. It is performed after seeding the producer cells and involves introducing the plasmid DNA into these cells to generate lentiviral particles. The success of this step is crucial for the subsequent stages of viral production, harvesting, and transduction. Our lentivirus packaging services are all performed according to the lentivirus packaging protocol submitted with the request. If you have concerns about your lentivirus transfection efficiency, contact our team, we are happy to review and optimize your protocol.
Let’s get started, our team is excited to bring your lentivirus research to life.
Learn about our quick turnaround lentivirus packaging services.
Want to learn more about the latest in lentivirus based research? Our colleagues at ScienceDirect and Genetic Engineering & Biotechnology News continuously collect and publish the latest information on lentivirus-based research.
Want to learn more about the latest in lentivirus based research? Our colleagues at ScienceDirect and Genetic Engineering & Biotechnology News continuously collect and publish the latest information.
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