Journal: iScience

Article Title: A key role for the exoribonuclease XRN1 in regulating the hepatitis B viral transcriptome

doi: 10.1016/j.isci.2026.116328

Figure Lengend Snippet: XRN1 regulation of the HBV transcriptome (A) HBV transcript mapping. Cartoon depicting the genomic viral RNA with the 5′ and 3′ epsilon stem loops, canonical transcription start sites, and major viral RNAs along with open reading frames (core, Pol, L, M, S, and X). HepG2-NTCP WT or XRN1 KO22 cells were infected with HBV, RNAs extracted at 6 dpi, and analyzed by long-read sequencing. Read counts for pC, pg, preS1, preS2, S, and X transcripts (left), along with spliced isoforms (right), are shown. Data are presented for 6 independent samples and statistical significance assessed using unpaired t tests (corrected for multiple comparisons, ∗ p < 0.05). (B) In vitro pgRNA digestion by XRN1. Capped-analog modified mRNA was synthesized by in vitro transcription. The capped and decapped pgRNA was incubated with XRN1, evaluated by agarose gel electrophoresis, and northern blotting using an HBV probe. Please see also .

Article Snippet: In vitro pgRNA transcription was performed in a 20 μL final volume using a T7 mScript standard mRNA production system by following the protocol provided by the manufacturer (CellScript), the DNA template was digested with 10 μL of a DNase cocktail containing 10 U of DNase I (NEB), 3 μL 10 X DNase I buffer, 2 μL nuclease-free water and incubated at 37°C for 20 min. After purification of the RNA using RNeasy kit (Qiagen), capping and polyadenylation were performed following Cap1 and Cap0 mRNA protocol described in the user’s manual, and the RNA was again purified using RNeasy kit.

Techniques: Infection, Sequencing, In Vitro, Modification, Synthesized, Incubation, Agarose Gel Electrophoresis, Northern Blot