Expression of proteins via a two-vector based inducible system
The invention provides a baculovius based expression System for targeted down-regulation of any gene on the bacuiovirusgenome or the cellular genome.
Products, such as proteins, VLPs and other nanoparticles are often produced in insect cells using the baculovirus system. However, purification of the final product is difficult and requires several tedious steps, because the baculovirusparticles are present as a DNA containing contaminant in the supernatant.
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Therefore, strategies have been tested to abolish baculovirus system secretion during the protein production process.
Usually, a gene that is essential for virus budding, is deleted from the baculovirus genome, and a helper cell line providing the missing gene is generated, that allows propagation of this virus. However, these helper cell lines suffer from instability and poor virus production. Antisense-RNA based strategies have been established in insect cell, usually targeting cellular genes. Most of these approaches are not efficient. The technology requires cell specific polymerase 1 promoters for exact transcription of the guide RNA. For identification of such promoters, the whole genome sequence must be available.
The invention provides a baculovirus based expression system for targeted downregulation of any gene on the baculovirus genome or the cellular genome. By antisense RNA or CRISPR/Cas9 technology, genes that are essential for baculovirus budding are being downregulated during the time of protein production. The downregulation mechanism is induced only when a combination of two different baculovirus species is present (dual vector system) and is regulated by the bacterial T7-System, which allows universal use in all insect cells as well as mammalian cells (no cell specific Polymerase 1 promotor required). The baculovirus species can be produced individually to high titers, only in combination, baculovirus production is hampered or abolished virus.
Application fields are in biotechnology medicine and pharma where high-quality proteins are used.
On behalf of University of Natural Resources and Life Sciences, Vienna TransMIT GmbH is looking for cooperation partners or licensees for distribution / further development in Germany, Europe, USA and Asia.
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Stichworteprotein expression, baculovirus expression system