Gene transduction in mammalian cells using Bombyx mori nucleopolyhedrovirus assisted by glycoprotein 64 of Autographa californica multiple nucleopolyhedrovirus.

ABSTRACT: Autographa californica multiple nucleopolyhedrovirus (AcMNPV), an alphabaculovirus, has been widely utilized for protein expression in not only insect cells but also mammalian cells. AcMNPV is closely related to Bombyx mori nucleopolyhedrovirus (BmNPV), and nucleotide sequences of AcMNPV genes have high similarity with those of BmNPV. However, the transduction of BmNPV into mammalian cells has not been reported. In this study, we constructed a recombinant BmNPV (BmNPV?bgp/AcGP64/EGFP) whose surface 64?kDa glycoprotein (BmGP64) was substituted with that from AcMNPV (AcGP64). BmNPV?bgp/AcGP64/EGFP also carried an EGFP gene under the control of the CMV promoter. BmNPV?bgp/AcGP64/EGFP successfully transduced HEK293T cells. In comparison, a control construct (BmNPV?bgp/BmGP64/EGFP) which possessed BmGP64 instead of AcGP64 did not express EGFP in HEK293T cells. The transduction efficiency of BmNPV?bgp/AcGP64/EGFP was lower than that of an AcMNPV based-BacMam GFP transduction control. This result indicates that AcGP64 facilitates BmNPV transduction into HEK293T cells. BmNPV can be prepared easily on a large scale because BmNPV can infect silkworm larvae without any special equipment, even though specific diet is needed for silkworm rearing. BmNPV gene transduction into mammalian cells can potentially be applied easily for gene delivery into mammalian cells.

SUBMITTER: Kato T 

PROVIDER: S-EPMC4999795 | biostudies-literature | 2016 Aug

REPOSITORIES: biostudies-literature