JCV miRNA sequence and detection.

Abstract

<p>(A) Schematic presentation of the JCV genome. The black circle marks the transcript location of the JCV miR-J1 stem loop. (B) JCV miR-J1-5p and -3p sequences are compared to the Merkel Cell Polyomavirus-, SV40- and BK virus-miRNA sequences. (C & D) CRC cells were transfected in vitro with a JCVT-Ag-E plasmid, and JCV T-Ag message and miRNA expression were analyzed. In C, GAPDH and β-actin were used as loading controls for mRNA and protein expression, respectively. (D) Vector transfected cells showed no detectable miR-J1-5p expression, while JCV miR-J1-5p expression was high in transfected cells. To measure the expression of miR-J1-5p, expression in the vector was set to a Ct-value of 40, and 2^−ΔΔCt values were calculated using RNU6b for normalization.</p