Fig. 3: Divergent qPCR primers selectively detect HIV circRNAs.
From: Backsplicing of the HIV-1 transcript generates multiple circRNAs to promote viral replication
a Divergent qPCR primer sets binding across either a BSJ or a canonical splice junction are utilized to amplify each HIV circRNA independently. b Total RNA was extracted from infected RevCEM-D4 cells and digested with RNase R or mock-digested as a control. Samples were amplified with the divergent circRNA qPCR primer sets and divergent primers (CirGP) for the linear Gag-Pol transcript as a negative control. Primers for the cellular gene RPL13A, all HIV-1 multiple spliced messengers (MS All, Fig.1) and all HIV-1 linear transcripts (HIV All, Fig. 1) were also utilized as controls. Data are represented as means ± SEM. P value (paired t-test) *<0.05, **<0.01, ***<0.001. No RT controls are shown in the Supplementary Source Data. c The relative percentage of each circular and linear RNA detected after RNase R digestion relative to the mock-digested sample is indicated.
