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. 2022 May 24;6(9):2474–2487. doi: 10.1002/hep4.2014

FIGURE 5.

FIGURE 5

Olaparib enhances the antiviral effect of HBV‐CRISPR in HBV‐infected hepatoma cells and PHHs. (A–C) HepG2‐hNTCP‐C4‐iCas9 cells transduced with HBV gRNA were treated with or without olaparib (1 μM) 10 days after HBV inoculation (10,000 GEq/cell). (A) Experimental protocol. (B) Intracellular cccDNA levels in HBV gRNA–transduced HepG2‐hNTCP‐C4‐iCas9 cells 13 days after HBV inoculation. (C) pgRNA levels in HBV gRNA–transduced HepG2‐hNTCP‐C4‐iCas9 cells 13 days after HBV inoculation. (D–F) HepG2‐hNTCP‐C4‐iCas9 cells transduced with HBV gRNA were treated with DOX 4 days after HBV inoculation (10,000 GEq/cell) and then with olaparib (1 μM) or vehicle 10 days after HBV inoculation. (D) Experimental protocol. (E) cccDNA levels in HBV gRNA–transduced HepG2‐hNTCP‐C4‐iCas9 cells 13 days after HBV inoculation (n = 4; *p < 0.05). (F) Intracellular pgRNA levels in HBV gRNA–transduced HepG2‐hNTCP‐C4‐iCas9 cells 13 days after HBV inoculation (n = 4; **p < 0.01). (G–I) PHHs isolated from humanized liver chimeric mice were lentivirally transduced with tandem Cas9‐expressing and HBV gRNA–expressing vectors (10 MOI) 19 days after HBV inoculation (500 GEq/cell) and were then treated with olaparib (1 μM) or vehicle. (G) Experimental protocol. (H) cccDNA levels in PHHs 35 days after HBV inoculation (n = 4; *p < 0.05). (I) Intracellular pgRNA levels in PHHs 35 days after HBV inoculation (n = 4, *p < 0.05).