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. Author manuscript; available in PMC: 2022 Jan 15.
Published in final edited form as: Virus Res. 2020 Nov 6;292:198224. doi: 10.1016/j.virusres.2020.198224

Figure 2. A plasmid with Cre recombinase in cisis stable and forms recombinant cccDNA molecules.

Figure 2.

(A) A CMV.NLS-Cre cassette, with inserted internal intron to inhibit bacterial expression, was cloned into the LoxP-HBV plasmids, such that the Cre transcript would utilize the HBV polyadenylation site. Upon introduction into mammalian cells, expressed Cre will excise the HBV genome forming to separate circular DNA molecules inside the cell. (B) The stability of the Cre/LoxP-HBV plasmids in E. coli were assessed by HindIII restriction digest, showing that their total size is the 7.9 kB of the original plasmid. (C) Upon transfection into 293T cells, recombination can be confirmed using PCR across the recombination junction showing predicted band sizes are present. (D) Livers of female NSG mice were transfected with 20 μg Cre/LoxP-HBV plasmids using hydrodynamic tail vein injection and HBsAg levels in the serum assessed week 1 post-injection (n=4). Data represent mean ± standard error of mean (SEM).