Applications of In-Vitro Transfection 

• Different genetically modified proteins 
• Drugs are produced through this technique. 
• Diagnosis 
• Treatment of diseases 
• Determination of efficacy of drugs 
• Suppressing the gene

Clinical Approach of In-Vitro Transfection 

Different molecular progress has been made, and trials using in-vitro transfection on the mammalian cells. Introducing genes as a therapy against various chronic diseases is possible due to this technology. The mitochondrial DNA mutations are easily modified by changing the mitochondrial DNA heteroplasmy in the cultured cells under control conditions. In this therapy, scientists harvest or isolate the cells from the patient and modify them in the laboratory, also known as ex vivo transfection. The cancer cell lines cultured cells help observe the different stages of cancer in-vitro and understand the physical structure, development, and growth rate of carcinoma. In the future 20 years, there is a possibility that the transfection technique will become a clinical part.  

If the in-vitro transfection, is the target organ is the heart, lung, or brain, so in-vivo transfection is more valuable than in-vitro due to efficacy level. 

RNA interference (RNAi) and small interfering RNA (siRNA) 

RNA interference (RNAi) is an efficient way to knock down certain genes and to observe the resulting changes in the phenotypes. This method is effective for identifying the phenotypes. Small interfering RNAs (siRNA) create an RNA-induced silencing compound (RISC) within the cell, which blocks the expression of the targeted gene.  

The most common methods used to deliver siRNA are lipid/polymer-mediated delivery and virus-mediated delivery. Despite the widespread application of siRNA, massive efforts are being made to create more efficient, secure, safe, and reliable methods for delivering siRNAs to cells due to the immense potential of RNAi for use in clinical trials to treat diseases. Two relatively new methods of transfection, including siRNA transfection and mRNA, are bringing innovative approaches to conduct cell research, each with their distinct advantages. 

siRNAs are transfected to achieve silencing of RNA (i.e., elimination of RNA and proteins from the target gene). This has been an effective method of research to obtain the “knockdown” of proteins of interest with the potential for application in the field of gene therapy. The limitations of the silencing method are the toxic nature of the transfection to cells and the potential “off-target” impacts on the production of other proteins or genes. 

Short and Long-RNA Transfection 

Short-RNA transfection is commonly used in biomedical research to shut down expression levels of a particular protein and to express or inhibit the activity of miRNA (using short RNA, which acts independently of the cell’s own RNAi machinery).  

It is not commonly referred to as siRNA). DNA-based vectors (viruses, viruses, and plasmids) that encode shorter RNA molecules may also be employed. Short-RNA transfection does not alter cells’ DNA, which is why short RNA is a very new type of macro-molecular drug.  

Long-RNA transfection is the method of intentionally inserting RNA molecules larger than approximately 25nm into a living cell. The difference between short and long RNA transfection is that long RNA molecules can trigger an immune system response in cells, which can lead to nonspecific reactions like cell cycle arrest and apoptosis.