Figure 6. Activation of A3C, A3D, and A3H by CRISPR and its
effects on HBV replication and cccDNA deamination. (A) Design
of gene-targeting CRISPRa sgRNA. (B) Activation of target genes
measured by qRT-PCR. (C) Anti-HBV activity of CRISPR-activated
genes determined by measuring pgRNA, cccDNA, and secreted HBsAg levels.
To further assess antiviral and deamination activity of A3C, A3D, and
A3H under HBV rcDNA-depleted conditions, we performed experiments with
anti-HBV siRNA (Figure 7). Combined treatment with siRNA, A3C, and A3H
enhanced reduction in HBV rcDNA levels and cccDNA levels. However, HBV
cccDNA deamination and secreted HBV DNA were not detected in either
group (Figure 7E, I). Similar to the results with A3A, A3B, and AID,
siRNA increased deamination of secreted HBV DNA with most factors
(Figure 7I).