How Long After Transfection GFP Expression (And Why)?

Exact Answer: 4 hours to 5 days

When some nucleic acids are artificially induced into healthy cells the process is termed transfection. The process allows nucleic acid to bypass the cell membrane to enter the cell. It is used in biological sciences to study various other altered cellular processes that result from the introduction of these RNA and DNA particles.

GFP, on the other hand, stands for green fluorescent protein. GFP is used as a reporting molecule for gene expression in biology since it emits green fluorescence under blue light. GFP expression is studied in regard to transfected cells. It is also used to study cell structures in embryos and fetuses.

How Long After Transfection GFP Expression?

Through the process of transfection, it becomes possible to introduce nucleic acids into mammalian cells without viral vectors. This gene transfer method allows researchers to study gene function and protein expression in cellular environments.

Generally, researchers note that in 90% of cells under observation, Green Fluorescent Protein is visible within 4 hours of transfection. However, in some cells, it may take up to 5 days to show the effects of transfection. In other words, in order for the green fluorescence protein to be visible under the microscope, it may take up to 5 days.

The GFP expression may be hinged on factors like the use of chemical or physical transfecting agents. In the case of the former, we find that GFP expression happens sooner rather than later. This would imply that the chemical transfection agent works faster on a cellular level than the physical agents.

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Alternatively, physical transfection agents may take a while to show GFP expression in the mammalian cells. They work slower than the rapid action chemical transfection agents. Moreover, researchers have also stated that the cell cycle stage, when a particular cell is selected also affects the time when GFP expression happens in the chosen cell.

It is equally important to note that research in this arena has been scanty. This is also a prime reason for the absence of extremely conclusive results on the issue. It is thus prudent to propose that the majority of mammalian cells will show GFP expression within 4 to 6 hours of the transfection.  

In Summary:

Transfection Agent UsedTime Taken to Show GFP Expression
Chemical Agents4 hours
Physical AgentsUp to 5 days

Why Does GFP Expression Take So Long After Transfection?

GFP expression is emblematic of the fact that the transfection has been successful and the gene structure now stands altered. For this to happen, the transfected agent has to be given time to initiate the structural alteration. Thus, it takes some time to be visible under a microscope.

GFP expressions may be rapid in some cells after transfection because of the kind of agent used to induce the nucleic acids into the mammalian cells. If a chemical vector is used, the result will invariably be hastened. One can then expect to see green fluorescent protein expression within a few hours of the transfection.

However, if physical agents are used, transfection expression through GFP will take some more time. Similarly, the cell structure may also delay the process by a few days. The cells have to be allowed to reach a level of stability before they can be tested for green fluorescent proteins under the microscope.

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The imbibition of the nucleic acids is bound to take a while because one is trying to manually alter the chemical composition of the cell. This is a significant structural alteration that cannot- in most cases- be streamlined by the use of a catalyst. Thus, as a scientist, you have to give the cells time to adapt and change at their own pace.

It is thus prudent to wait for up to a week to see if the GFP expression is visible in cell clusters. Since different cells react differently, time differences in gene expression are only natural and expected.

Conclusion

Transfection can be a very novel and useful way of changing a cell’s chemical composition and gene expression. It is often used by scientists to study mammalian cells and their response to the induced nucleic acids.

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Generally, cell clusters tend to demonstrate a successful transfection session within as early as 4 to 6 hours of the process. This is expressed through the green fluorescent protein that is visible in the cells under a microscope. In very few cases, cells may take longer to show the changed gene expression. This time period can extend up to 5 days in such rare events.  

References

  1. https://link.springer.com/article/10.1007/s004240050445
  2. https://www.sciencedirect.com/science/article/pii/S0168010298001138