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).