key term - 293T Cells

Definition

293T cells are a human embryonic kidney cell line that has been genetically modified to express the SV40 large T antigen, making them highly efficient for transfection and protein expression studies. This cell line is widely used in laboratories for various applications, including the production of recombinant proteins and viral vectors due to their ease of culture and manipulation.

5 Must Know Facts For Your Next Test

1. 293T cells are derived from the original 293 cell line, which was established from human embryonic kidney cells in the early 1970s.
2. The expression of SV40 large T antigen in 293T cells allows for enhanced replication of transfected plasmids, making them ideal for producing viral vectors.
3. These cells can grow in suspension or adherent cultures, providing flexibility for various experimental setups.
4. 293T cells are commonly used in gene therapy research due to their ability to produce high yields of viral vectors for delivering therapeutic genes.
5. They are also frequently utilized in studies involving protein-protein interactions and post-translational modifications.

Review Questions

• How do 293T cells enhance the efficiency of transfection compared to other cell lines?
  • 293T cells enhance transfection efficiency due to their high expression levels of SV40 large T antigen, which promotes the replication of transfected plasmids. This makes them particularly effective for experiments requiring high levels of protein expression or viral vector production. The ability to maintain high viability and growth rates under laboratory conditions also contributes to their popularity as a host for transfection experiments.
• Discuss the role of SV40 large T antigen in the functionality of 293T cells and its implications for research.
  • SV40 large T antigen plays a crucial role in the functionality of 293T cells by allowing for the replication of plasmids that contain origins of replication recognized by SV40. This feature enables researchers to achieve higher yields of recombinant proteins or viral vectors compared to other cell lines. Additionally, this characteristic has implications for gene therapy applications, as it allows for efficient production of viral vectors necessary for delivering therapeutic genes into target cells.
• Evaluate the advantages and limitations of using 293T cells for producing recombinant proteins and viral vectors in research.
  • Using 293T cells presents several advantages, including high transfection efficiency, rapid growth rates, and adaptability to both suspension and adherent cultures, which facilitate large-scale protein or viral vector production. However, there are limitations such as potential post-translational modifications that may differ from those in native human cells, which could affect the functionality of produced proteins. Additionally, depending on the application, researchers must be cautious about using transformed cell lines like 293T due to potential differences in biological behavior compared to primary cells.

Related terms

Transfection: The process of introducing nucleic acids into cells to study gene function and protein expression.

SV40: Simian Virus 40, a virus that can induce tumors in animals and is used in research to drive the expression of genes in transformed cells.

Recombinant Protein: Proteins that are artificially made through the combination of DNA from different sources, often produced using cell lines like 293T.