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. 2021 Jun 16;12:696604. doi: 10.3389/fmicb.2021.696604

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Copyright © 2021 Shao, Shen, Wang and Qiu.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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rAAV2/HBoV1 vector production systems. (A) NS-free rAAV2/HBoV1 vector production system in HEK293 cells. pAAV2 carries the genes of interest in rAAV2 genome. pCMVoptVP1-AAV2Rep is a two-in-one plasmid, in which the VP1 and Rep2 (Rep78/52) are expressed in two independent cassettes. pAd4.1-CMVoptVP2/3 has the optimized (opt)VP2/3 expression cassette in the Ad helper gene expression plasmid pAd4.1. (B) rAAV2/HBoV1 baculovirus expression vector (BEV) production system. Bac-AAV2ITR-GFP-Luc carries an rAAV2 genome. Bac-AAV2Rep-HBoV1Cap expresses AAV2 Rep proteins and HBoV1 capsid proteins. Bac-HBoV1NP1 expresses HBoV1 NP1. P10 and Ph are baculoviral promoters, and CMV and F5 tg83 are a human cytomegalovirus major immediate-early promoter and a synthetic promoter, respectively. Various polyadenylation signals are indicated as polyA. Luc: firefly luciferase gene; GFP, enhanced green fluorescent protein gene, and mCherry, a monomeric red fluorescent protein gene. They can be replaced by any gene of interest (GOI) for expression in the vector.