Event Title

Identification of Potential Viral Circular RNA in Epstein Barr Virus-Producing Cells

Date

12-2015 12:00 AM

Major

Nutritional Science, Microbiology

Department

Food Science and Human Nutrition

College

College of Human Sciences

Project Advisor

Luwen Zhang; Kevin Schalinske

Project Advisor's Department

Nebraska Center for Virology, University of Nebraska–Lincoln (Zhang); Food Science and Human Nutrition (Schalinske)

Description

Circular RNAs (circRNAs) are a recently-discovered class of RNA molecules with a unique closed-loop structure. While circRNAs may be almost exclusively non-coding, several regulatory roles have been proposed. The role of circRNAs in disease, particularly cancer, is under investigation. Prior to this study, a gene in the Epstein Barr Virus (EBV) genome, BHLF1, was identified as having a putative circRNA. The purpose of this study was to identify whether BHLF1 RNA is circular and to determine how expression of BHLF1 RNA varies with time and cell media conditions. To this end, BHLF1 expression was induced in Akata cells (a Burkitt’s lymphoma cell line which produces transforming EBV) under different media conditions and cells were collected at various time points. RNA was extracted and cDNA was synthesized using RT-PCR. A product of the correct size, 283 bp, was successfully amplified using outward facing BHLF1 primers, indicating that the original RNA was circular. Expression of this product depended on time of collection of Akata cells and media conditions. Because circRNAs have been shown to have roles in gene regulation and disease development, identification of BHLF1 RNA as circRNA may offer a new insight into its role in EBV pathogenesis.

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Dec 1st, 12:00 AM

Identification of Potential Viral Circular RNA in Epstein Barr Virus-Producing Cells

Circular RNAs (circRNAs) are a recently-discovered class of RNA molecules with a unique closed-loop structure. While circRNAs may be almost exclusively non-coding, several regulatory roles have been proposed. The role of circRNAs in disease, particularly cancer, is under investigation. Prior to this study, a gene in the Epstein Barr Virus (EBV) genome, BHLF1, was identified as having a putative circRNA. The purpose of this study was to identify whether BHLF1 RNA is circular and to determine how expression of BHLF1 RNA varies with time and cell media conditions. To this end, BHLF1 expression was induced in Akata cells (a Burkitt’s lymphoma cell line which produces transforming EBV) under different media conditions and cells were collected at various time points. RNA was extracted and cDNA was synthesized using RT-PCR. A product of the correct size, 283 bp, was successfully amplified using outward facing BHLF1 primers, indicating that the original RNA was circular. Expression of this product depended on time of collection of Akata cells and media conditions. Because circRNAs have been shown to have roles in gene regulation and disease development, identification of BHLF1 RNA as circRNA may offer a new insight into its role in EBV pathogenesis.