2021
DOI: 10.2174/1566523221666210118111657
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Popularizing Recombinant Baculovirus-derived OneBac System for Laboratory Production of all Recombinant Adeno-associated Virus Vector Serotypes

Abstract: Objective: On the basis of our previously established single recombinant baculovirus expression vector (BEV)-derived OneBac system, we have optimized the process and expanded the rAAV production range to the full range of serotypes rAAV1-13. Firstly, the AAV Cap gene was optimized to translate by ribosome leaky scanning and the gene of interest (GOI) was cloned into the pFD/Cap-(ITR-GOI)-Rep2 shutte plasmid. Following the classical Bac-to-Bac method, sufficient BEV stock containing all rAAV packaging elements… Show more

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Cited by 9 publications
(3 citation statements)
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“…We have previously established a single baculovirus-insect cell-based large-scale production system (OneBac system) and verified the preparation of rAAV1-13 serotypes [28, 29]. Although it is conducive to large-scale production, putting all the genetic elements in the common bacmid is not convenient for switching the carried foreign genes.…”
Section: Resultsmentioning
confidence: 99%
“…We have previously established a single baculovirus-insect cell-based large-scale production system (OneBac system) and verified the preparation of rAAV1-13 serotypes [28, 29]. Although it is conducive to large-scale production, putting all the genetic elements in the common bacmid is not convenient for switching the carried foreign genes.…”
Section: Resultsmentioning
confidence: 99%
“…We previously established a single baculovirus–insect cell system for large-scale virus production (OneBac system) and successfully used this system for the preparation of AAV1-13 serotypes [ 15 ]. To find AAV serotypes more suitable for precise in situ targeting, we screened AAV serotypes that were not commonly used for gene delivery purposes and found that AAV13 exhibited confined diffusion without retrograde infection and trans-synaptic transmission in the brain, and that its local diffusion range was smaller than AAV2.…”
Section: Introductionmentioning
confidence: 99%
“…AAV5, AAV8(Thomas et al, 2009), and AAV9(Kang et al, 2009).4.1.3 | Recombinant baculovirus infection of Sf9 (rBV-Sf9)Similar to rHSV-HEK293 systems, in rBV infection, Sf9 cells are infected with two or three rBVs carrying GOI, rep, and cap but no helper virus is required. rBV-Sf9 has been applied to rAAV serotyped 1-13 for production(Wu et al, 2021). Infection with separate rep and cap vectors allows better control of gene expression levels.…”
mentioning
confidence: 99%