BacMam technologies in cells and animals; Advances towards the transport of DNA in mammals

SIMONIN ALEJANDRO; OLEA FERNANDA DANIELA; BELAICH MARIANO NICOLÁS; GIMÉNEZ, CARLOS SEBASTIÁN; CROTTOGINI ALBERTO; NÚÑEZ PEDROZO CRISTIAN NAHUEL; GHIRINGHELLI D

Valencia

Congreso; International Congress on Invertebrate Pathology and Microbial Control & 52nd Annual Meeting of the Society for Invertebrate Pathology & 17th Meeting of the IOBC‐WPRS Working Group ?Microbial and Nematode Control of Invertebrate Pests.; 2019

Baculoviruses are insect viruses that efficiently transduce mammalian cells, remaining as non-replicative episomes. In this context, we set out to develop and evaluate recombinant AcMNPV, both in vitro and in animal models. Thus, 3 viruses were generated that express a fluorescent protein (eGFP, mCherry or BFP) under the CMV viral promoter. Through their use, transduction efficiency was optimized and measured on mammalian cells, which were analysed by fluorescence microscopy and flow cytometry. Using such results, in vivo tests were then performed on rabbits with peripheral artery ischemia (PAD). These tests revealed an adequate performance in short times (less than 7 days). Besides, another virus was generated that expresses a mutant version of HIF gene (Hypoxia inducible factor), with the purpose of proving its usefulness in a therapy for PAD. In parallel, to lengthen the expression of transgenes in vivo, a virus containing a gene circuit that allows the release of a minicircle by Cre/LoxP technology in an inducible manner (TetON/TetOFF system), is being developed. This tool will allow a controllable and lasting expression of the gene of interest. So far, we have preliminary in vitro results, which have shown a correct functioning. In summary, evaluations carried out in vitro and in vivo using recombinant baculoviruses showed satisfactory results, which are necessary to proceed with the preclinical evaluations of gene therapy active ingredients.