Key Difference – Transient vs Stable Transfection
Transfection is a process which is involved in the gene transfer of eukaryotic cells using chemical or physical methods. Transfection can be classified into main two types named transient transfection and stable transfection. During transient transfection, the gene of interest fails to integrate with the host genome and is expressed temporally within host for a short term whereas, in stable transfection, the gene of interest integrates with the host genome and is sustained for a long term in several generations. This is the key difference between transient and stable transfection. In both cases, transfection is successful, and genes are expressed.
CONTENTS
1. Overview and Key Difference
2. What is Transient Transfection
3. What is Stable Transfection
4. Side by Side Comparison – Transient vs Stable Transfection
5. Summary
What is Transient Transfection?
Transfection is an important tool for inserting genes into eukaryotic cells. Among the two types of transfection, transient transfection is a common way in gene transfer. Through a vector, foreign genes are transformed inside host cells. Once foreign DNA enters the host cell, it has two options. It can either integrate with host genome and replicate, or stay inside without integrating into the genome. Transient transfection exhibits the temporary expression of the inserted genes without integrating into the host genome. Genes express and produce the coded protein till the cell divides. However, due to the inability of integration, it cannot replicate and enter into future generations. This type of transfection is successful for a short time period. During the cell division or due to some other factors, foreign DNA is subjected into degradation. Transient transfection is shown when the foreign DNA is in the form of highly coiled DNA.
Figure 01: Transient Transfection
What is Stable Transfection?
Stable transfection shows successful foreign gene integration into host genome. Once foreign DNA enters inside the host cell, part of the foreign DNA integrates with the host genome and become a part of it. Hence, foreign DNA also replicates and passes into future generations when the host genome replicates. This type of transfection is complex and rare. However, due to the stable transfection into the genome, this trait is sustained for a longer period of several generations.
Stable transfection is a difficult process and requires an effective delivery of DNA and cell acquisition of the foreign DNA into its genome. Hence, linear DNA favors the stable transfection than circular DNA. However, stable transfection rate is approximately one in 104 transformed cells. Stable transfection can be observed by the cotransfomation of a selectable marker and performing the artificial selection on a medium.
Figure 02: Stable Transfection
What is the difference between Transient and Stable Transfection?
Transient vs Stable Transfection |
|
| Foreign DNA does not integrate into a host genome. | Foreign DNA integrates with host genome and becomes a part of it. |
| Replication within the Host | |
| Transiently transfected genes do not replicate within the host. Therefore genes are not passed to next generations. | Genes are replicated within the host and passed to future generations. |
| Time duration of the Gene Expression | |
| Genes are expressed for a finite time period and after that, they are destroyed. | Genes become part of the genome and expressed within the generation for a long time period. |
| Use | |
| This is useful for studying the effects of short-term expression of genes or gene products. | This is useful for studying the effects of gene expressions long term. |
| Detection of the Transfection | |
| Genes are expressed and can be easily detected by inserting a reporter gene. | Stable transfection can be easily detected by inserting a selectable marker and selecting through artificial selection on media. |
| Occurrence and Process | |
| Transient transfection is common and not complex to perform. | Stable transfection is rare in occurrence and complex to perform. |
| Nature of DNA | |
| Highly supercoiled DNA are suitable for transient transfection. | Linear DNA are suitable for stable transfection. |
| Applications | |
| This is used for studies of gene knockdown or silencing with inhibitory RNAs, protein production on a small scale | This is used for protein production on a large-scale, longer-term pharmacology studies, gene therapy, research on the mechanisms of long-term genetic regulation |
Summary – Transient vs Stable Transfection
Transient and stable transfections are two types of transfections shown during the gene transferring into eukaryotic cells by non-viral based systems. Transformed foreign DNA is not integrated with host genome in transient transfection while it integrates with host genome in stable transfection. Circular coiled DNA is shown transient transfection whereas linear DNA prefers stable transfection. This is the difference between transient and stable transfection. However transient transfection is more common and easy compared to stable transfection. But the selection of one among two depend on the purpose of the gene transfer and duration of the research project.
References:
1. Condreay, J. Patrick, Sam M. Witherspoon, William C. Clay, and Thomas A. Kost. “Transient and stable gene expression in mammalian cells transduced with a recombinant baculovirus vector.” Proceedings of the National Academy of Sciences of the United States of America. The National Academy of Sciences, 05 Jan. 1999. Web. 25 Mar. 2017
2. Kim, Tae Kyung, and James H. Eberwine. “Mammalian cell transfection: the present and the future.” Analytical and Bioanalytical Chemistry. Springer-Verlag, Aug. 2010. Web. 25 Mar. 2017