CRISPR activation of A3С, A3D, and A3H suppresses HBV
replication but does not affect cccDNA or the host genome.
As we showed, short-term activation of A3A and A3B effectively
suppresses HBV replication but causes off-target deamination of the host
genome. Therefore, we decided to analyze the antiviral efficacy of other
members of the APOBEC3/AID family with previously reported anti-HBV
activity (A3C, A3D, and A3H). We designed sgRNAs targeting promoters of
the selected genes (Figure 6A) and analyzed the effects of CRISPRa of
these genes in the HepG2 rcccDNA co-transfection model. Transcription ofA3C , A3D , and A3H was increased 2–20-fold (Figure
6B). HBV life cycle analysis revealed a 50% reduction in both pgRNA and
cccDNA levels following CRISPRa, while HBsAg level remained unchanged
(Figure 6C).