Differential Expression Pattern of linc-ROR Spliced Variants in Pluripotent and Non-Pluripotent Cell Lines.

OBJECTIVE

The human large intergenic non-coding RNA-regulator of reprogramming program () is known as a stem cell specific linc-RNA. linc-ROR counteracts differentiation via sequestering microRNA-145 (miR-145) that targets OCT4 transcript. Despite the research on the expression and function, the exact structure of transcripts is not clear. Considering the contribution of alternative splicing in transcripts structures and function, identifying different spliced variants of is necessary for further functional analyses. We aimed to find the alternatively spliced transcripts of and investigate their expression pattern in stem and cancer cell lines and during neural differentiation of NT2 cells as a model for understanding linc-ROR role in stem cell and differentiation.

MATERIALS AND METHODS

In this experimental study, locus was scanned for identifying novel exons. Different primer sets were used to detect new spliced variants by reverse transcription polymerase chain reaction (RT-PCR) and direct sequencing. Quantitative PCR (qPCR) and RT-PCR were employed to profile expression of transcripts in different cell lines and during neural differentiation of stem cells.

RESULTS

We could discover 13 novel spliced variants of linc-ROR harboring unique array of exons. Our work uncovered six novel exons, some of which were the product of exonized transposable elements. Monitoring expression profile of the spliced variants in a panel of pluripotent and non-pluripotent cells exhibited that all transcripts were primarily expressed in pluripotent cells. Moreover, the examined spliced variants showed a similar downregulation during neural differentiation of NT2 cells.

CONCLUSION

Altogether, our data showed despite the difference in the structure and composition of exons, various spliced variants of showed similar expression pattern in stem cells and through differentiation.