Baculovirus transactivator IE1 is functional in mammalian cells.

Murges, Daniela; Kremer, Andreas; Knebel-Mörsdorf, Dagmar

doi:10.1099/0022-1317-78-6-1507

Cited by 31 publications

(29 citation statements)

References 29 publications

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“…These results highlight the cell dependence of AAD gene activation and the promiscuous nature of the AcMNPV IE1 AAD. Previous studies have shown that the native AcMNPV IE1 is able to activate gene expression in mammalian BHK-21 cells (Carbonell et al, 1985;Carbonell & Miller, 1987;Murges et al, 1997). Our results indicate that this is due, at least in part, to the AAD domain of IE1, which appears to be a more universal activation domain, similar to VP16 AAD.…”

Section: Discussionsupporting

confidence: 59%

The acidic activation domains of the baculovirus transactivators IE1 and IE0 are functional for transcriptional activation in both insect and mammalian cells

Dai

Willis

Huijskens

et al. 2004

Journal of General Virology

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The acidic activation domains (AADs) of the baculovirus transactivators IE1 and IE0 are essential for transcriptional transactivation. To compare the relative transcriptional activation potentials of IE1 and IE0 AADs of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) and Orgyia pseudotsugata MNPV (OpMNPV), we constructed two ecdysone receptor (EcR)-based inducible expression systems to analyse six baculovirus AADs in two insect cell lines (Ld652Y and Sf9) and two mammalian cell lines (NIH-3T3 and CHO). For insect cell expression, the AADs were fused to the C, D, E and F domains of the spruce budworm Choristoneura fumiferana EcR. For mammalian cell expression the AADs were fused to the E and F domains of mammalian Mus musculus retinoid X receptor. In Ld652Y and Sf9 cells, chimeric proteins containing the AcMNPV AADs activated gene expression to higher levels than those containing the OpMNPV AADs. In NIH-3T3 cells, chimeras containing AcMNPV IE1 and IE0 AADs consistently activated gene expression to higher levels than the archetypal mammalian herpesvirus VP16 AAD. In contrast, OpMNPV AADs only activated expression by 5-15 % relative to the VP16 AAD. In CHO cells, both AcMNPV and OpMNPV AADs exhibited intermediate transactivation levels relative to VP16 AAD. These results show that the baculovirus AADs are functional for transcriptional activation in mammalian cells and that AcMNPV AADs generally appear to be more potent than OpMNPV AADs in both insect and mammalian cells.

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Section: Discussionsupporting

confidence: 59%

The acidic activation domains of the baculovirus transactivators IE1 and IE0 are functional for transcriptional activation in both insect and mammalian cells

Dai

Willis

Huijskens

et al. 2004

Journal of General Virology

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“…The most successful use of baculovirus for delivery and expression of genes in mammalian cells used a CMV promoter and transduction at 37OC (10)(11)(12)(13)(14)(15)(16)(17). We found that transduction of primary pancreatic dissociated islet cells and other cell lines using baculovirus at 37OC was four-to fivefold higher than that at room temperature (L.M., L.H.P., unpublished data).…”

Section: Discussionmentioning

confidence: 98%

“…Recently, it was shown that baculovirus can be transported into some primary cells and into many cell lines derived from mouse, rat, porcine, and human tissues (10)(11)(12)(13)(14)(15)(16)(17). Using mammalian promoters, baculovirus enabled efficient transient expression of reporter genes in mammalian cells (10)(11)(12)(13)(14)(15)(16)(17). Condreay et al (17) also reported stable baculovirus transduction of several mammalian cell types with expression of a reporter gene for multiple passages.…”

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confidence: 99%

Baculovirus-mediated gene transfer into pancreatic islet cells.

Tamarina

Wang

et al. 2000

Diabetes

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Baculovirus transduction is a gene transfer method that uses a moth cell virus for mammalian cells in culture, which results in a high-level prolonged expression. Here we demonstrate that recombinant baculoviruses can serve as efficient gene transfer vehicles for delivering foreign genes driven by mammalian promoters into human and mouse pancreatic islet cells. Existing methods, such as various transfection and electroporation techniques, either suffer from low efficiency or cause extensive membrane damage. Viral vectors have emerged as an important tool for gene delivery and expression in mammalian cells but suffer from several drawbacks, such as lengthy construction time and expression of viral genes. The baculovirus Autographa californica multiple nuclear polyhedrosis virus is widely used as a vector for expression of foreign genes in insect cells and, more recently, in some mammalian cells. Using several green fluorescent protein- and LacZ-expressing constructs in a cytomegalovirus promoter cassette, we obtained efficient gene expression in primary human and mouse islet cells. There was no impairment of glucose-stimulated intracellular free calcium responses after baculovirus infection. The safety and the relative ease of construction and propagation of the virus makes the baculovirus system a useful tool for facilitating the transfer of foreign genes.

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“…The persistence of large regions of the viral genome in cells, which we have demonstrated in this report, raises the possibility that immediate early viral gene products could be expressed in mammalian cells that receive a virus. One of these gene products, the IE-1 transactivator, has been demonstrated to activate transcription from viral early promoters in mammalian cells (20).…”

Section: Discussionmentioning

confidence: 99%