Skip to main content
. Author manuscript; available in PMC: 2016 Jan 31.
Published in final edited form as: Virology. 2014 Dec 29;476:196–205. doi: 10.1016/j.virol.2014.12.001

Figure 2. Suppression HBV replication by HBV-specific Cas9/sgRNA combinations.

Figure 2

HBV replication was first induced in the HepAD38 cells by culture in the absence of Tet for 48 hrs. Then, the cells were transduced with lentiviral vectors encoding Spy Cas9/sgRNA combinations specific for the HBV RT, surface Ag or core ORFs, puromycin selected, and cultured in the continued absence of Tet. HepAD38 cells were also maintained in Tet+ media as a negative control. The HepAD38 cells and supernatant media were harvested and total HBV DNA (A) and intracellular HBV DNA or cccDNA (B) quantified by qPCR after 10 or 14 days in culture, respectively. (+) indicates the sample was below the detection limit (> 45 cycles). All results of qPCR assays were normalized to the N.S. sgRNA control cell line. Levels of HBsAg (C) and HBeAg (D) secreted into the culture media were measured by ELISA after 10 days in culture. Data are displayed as the mean ± SD of replicates. Statistical analyses were performed using Student’s t-test for comparison between two groups using JMP pro 10 software. A value of P < 0.05 (*) was considered statistically significant. (***) represents P < 0.001.