The 3'UTR region of the SLC22A2 gene cloned into the psiCHECK vector exhibited significantly increased expression in HEK293T cells in the presence of miR-H3 of HSV-1
The 3'UTR region of the SLC22A2 gene cloned into the psiCHECK vector exhibited significantly increased expression in HEK293T cells in the presence of miR-H3 of HSV-1
Introduction: Herpes simplex virus type 1 (HSV-1) is a common viral infection known primarily for causing oral and labial herpes, often referred to as cold sores.. If HSV-1 spreads to the brain and central nervous system, serious infections such as herpes simplex encephalitis may occur, leading to irreversible complications. "This virus can establish colonization within the body during childhood and persist throughout a significant portion of the population's lifetime without any disease manifestation. As the virus depends on the host cell's machinery, a bidirectional interaction with the host must occur during latency or even during infection itself. One of the primary regions where the virus resides is within the ganglia of the nervous system. The ion channels, among the essential components of the nervous system, play a crucial role in brain homeostasis. HSV-1, similar to humans, produces a number of non-coding RNA molecules known as miRNAs that impact the expression of a wide range of host genes. Following bioinformatic studies and Real-time PCR analysis, it was determined that the SLC22A2 or OCT2 gene, which belongs to the category of cation transporters, exhibits significant upregulation by the virus's miR-H3 (3P).
Methods: For the 3'UTR region of the SLC22A2 gene, we designed primers and, after amplifying it and performing transformation, cloned the fragment into the psiCHECK vector. Then, we transfected this vector into HEK293 cells in the presence of miR-H3. After extracting RNA and performing real-time PCR, we observed that in the presence of miR-H3, there was an increase in expression. To confirm this, we used a luciferase assay, which showed an expression increase of up to 80% in the presence of miR-H3.
Results: The SLC22A2 gene has been recognized in recent years as a drug transporter. Additionally, it was first identified in 2017 that dysfunction of this gene plays a role in neurological diseases such as epilepsy . Given that the HSV-1 virus can lead to acute conditions such as epileptic encephalopathy, this gene could serve as a suitable therapeutic target for disease targeting.
After extracting RNA and performing real-time PCR, we observed that in the presence of miR-H3, there was an increase in expression. To confirm this, we used a luciferase assay, which showed an expression increase of up to 80% in the presence of miR-H3.
Conclusion: For the 3'UTR region of the SLC22A2 gene, we designed primers and, after amplifying it and performing transformation, cloned the fragment into the psiCHECK vector. Then, we transfected this vector into HEK293 cells in the presence of miR-H3. After extracting RNA and performing real-time PCR, we observed that in the presence of miR-H3, there was an increase in expression. To confirm this, we used a luciferase assay, which showed an expression increase of up to 80% in the presence of miR-H3.