What is transient overexpression? When it comes to the development of scientific knowledge and research, especially cell biology, transient overexpression techniques have been perpetually utilized for decades.
The vast majority of popular studies that make use of cell signaling have in most cases used transient overexpression in one way or another. However, recently these methods have come into question, with some scientists pushing for a change in how cell signaling research is conducted and studied. Let us delve more deeply into the significance of transient overexpression, and its significance in the research of cell biology.
Transient overexpression, also referred to as ‘transient gene expression’ is a short-term appearance or expression of genes. These genes become altered after nucleic acids are introduced into the eukaryotic cells. A chemical agent known as calcium phosphate is used as a means of transportation into the eukaryotic cells in a process called transfection. Furthermore, transient overexpression is uniquely different to the scientific methods known as ‘stable expression’. A significant distinguishing feature is that the vector carrying the DNA does not directly merge with the hosting DNA cell, and because of this the vector inevitably disappears after numerous replication cycles.
Most scientific studies that make use of transient overexpression tend to utilize cultured animal cells. In addition, the same technique is also applied in the study and research of plant cells. The difference in the two methods lays in how the nucleic acids travel from one place to another.
Nonetheless, whether it be plant research, or the study of animals the usage of transient overexpression should cause a transient transfer of nucleic acids. If the method was to cause a long-term transfer rather than a transient one it would have to be classified as ‘stable expression’.
Understanding the significance of Transient Overexpression in Research?
Transient overexpression plays an important role in the advancement of scientific research. For example, transient overexpression is frequently utilized in the study of point mutations often leading to truncations of certain proteins. The absence of a particular function due to the effects of mutations is often regarded as evidence for the function being inactivated.
Additionally, although this can be considered to be demonstrative evidence, the chance still exists that other factors could have contributed to the inactivated function. For example, the accumulation of a distorted protein in the same environment of transient overexpression can create a situation in which the data is deceptive.
Furthermore, another significant aspect of transient overexpression is the possibility of being able to characterize the effect of abnormal protein mutations. Overexpression research is highly regarded because of the potential to rescue phenotypes that can be seen in what is known as a loss of function model. Scientists have found that the insertion of a mRNA coding sequence can be utilized to create transient overexpression in certain animals. Most recently, zebrafish have been made use of in the study and advancement of overexpression.
The utilization of zebrafish in the study of transient overexpression
Zebrafish have proven to be especially useful in the study as well as research of transient overexpression. Zebrafish have a genetic composition that is similar to human beings.
In a typical scientific study, one cell stage embryos of zebrafish would be taken and inserted with a solution which is comprised of a synthetic mRNA. This is also known as a Morpholino oligomer. . These studies show how the transient overexpression of injected embryos can be readily identified at an early stage. Applications of these studies include the following:
- The assessment of mRNA toxicity at various levels.
- The protection of phenotypes as they pertain to genes as well as loss of function.
- Temporary state of transient expression.
- More affordable checks of genes whose expression is supposed to take on a pathological role.
Apart from these morpholinos or antisense oligonucleotides injected in embryos, there are also other techniques performed at Biobide that use CRISPR-mediated mutants called Crispants. These consist of a sequential transfection expression system that also enables transient expression of proteins.
Without a doubt as the studies of transient overexpression continue, the groundbreaking discoveries will prove to be extremely beneficial for humanity as whole.
