Autoregulated, bidirectional and multicistronic gas-inducible mammalian as well as lentiviral expression vectors

Hartenbach, Shizuka; Fussenegger, Martin

doi:10.1016/j.jbiotec.2005.03.025

Cited by 22 publications

(31 citation statements)

References 41 publications

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“…compounds (e.g., tetracycline, Prati et al, 2002;streptogramins, Moser et al, 2000;macrolides, Weber et al, 2002;gaseous acetaldehyde, Hartenbach and Fussenegger, 2005); (ii) autoregulated (Fussenegger et al, 1997;Hartenbach and Fussenegger, 2005); (iii) dual-regulated ); (iv) delivered using retroviral or lentiviral configurations (Fux et al, 2004a;Hartenbach and Fussenegger, 2005;Mitta et al, 2002); and (v) delivered on ever more compact and efficient genetic platforms (Fux et al, 2004a). To date, however, these vectors have only facilitated expression of multiple transgenes, and not downregulation of target genes.…”

Section: Discussionmentioning

confidence: 99%

Multi‐gene engineering: Simultaneous expression and knockdown of six genes off a single platform

Greber

Fussenegger

2006

Biotech & Bioengineering

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Increases in our understanding of gene function have greatly expanded the repertoire of possible genetic interventions at our disposal with the consequence that many genetic engineering applications require multiple manipulations in which target genes can be both overexpressed and silenced in a simple and co-ordinated manner. Using synthetic introns as a source of encoding shortinterfering RNA (siRNA), we demonstrate that it is possible to simultaneously express both a transgene and siRNA from a single polymerase (Pol) II promoter. By encoding siRNA as an intron between two protein domains requiring successful splicing for functionality, it was possible to demonstrate that splicing was occurring, that the coding genes (exonic transgenes) resulted in functional protein, and that the spliced siRNA-containing lariat was capable of modulating expression of a separate target gene. We subsequently extended this concept to develop pTRIDENT-based multi-cistronic vectors that were capable of co-ordinated expression of up to three siRNAs and three transgenes off a single genetic platform. Such multi-gene engineering technology, enabling concomitant transgene overexpression and target gene knockdown, should be useful for therapeutic, biopharmaceutical production, and basic research applications.

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Section: Discussionmentioning

confidence: 99%

Multi‐gene engineering: Simultaneous expression and knockdown of six genes off a single platform

Greber

Fussenegger

2006

Biotech & Bioengineering

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“…A detailed description of plasmid construction is shown in supporting information (SI) Text. pWW993 (P ETR -ADH-pA), erythromycin-regulated ADH (7, 13); pWW926 (P hEF1␣ -BTDpA), constitutive human biotinidase (BTD); pWW811 (P SV40 -ES-pA), erythromycin repressor E fused to streptavidin (ES) (13,14); pWW1015 (P AIR -sBLA-pA), acetaldehyde-inducible sBLA (17,21); pWW1019 (P hCMV -sBLA-pA), constitutive sBLA (22); pWW1016 (P AIR -NEO-pA), acetaldehyde-inducible neomycin resistance; and pWW1018 (P AIR -RipDD-pA), acetaldehyde-inducible RipDD (14,23).…”

Section: Methodsmentioning

confidence: 99%

Synthetic ecosystems based on airborne inter- and intrakingdom communication

Weber

Baba

Fussenegger

2007

Proc. Natl. Acad. Sci. U.S.A.

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Intercellular communication within an organism, between populations, or across species and kingdoms forms the basis of many ecosystems in which organisms coexist through symbiotic, parasitic, or predator-prey relationships. Using multistep airborne communication and signal transduction, we present synthetic ecosystems within a mammalian cell population, in mice, or across species and kingdoms. Inter-and intrakingdom communication was enabled by using sender cells that produce volatile aldehydes, small vitamin-derived molecules, or antibiotics that diffuse, by gas or liquid phase, to receiver cells and induce the expression of specific target genes. Intercellular and cross-kingdom communication was shown to enable quorum sensing between and among mammalian cells, bacteria, yeast, and plants, resulting in precise spatiotemporal control of IFN-␤ production. Interconnection of bacterial, yeast, and mammalian cell signaling enabled the construction of multistep signal transduction and processing networks as well as the design of synthetic ecosystems that mimic fundamental coexistence patterns in nature, including symbiosis, parasitism, and oscillating predator-prey interactions.biological circuit ͉ gene switch ͉ synthetic biology I ntercellular cross-talk either within or between organisms of the same or different species represents a fundamental communication process that is responsible not only for orchestrating vital functions within multicellular life forms but also for determining the manner in which different organisms coexist. Whereas the most sophisticated intercellular communication networks manage processes such as cellular differentiation and pattern formation during development or use hormonal and neural circuitries to adapt responses by individual organisms to endogenous and exogenous stimuli, higher-order ecosystems are organized by basic interaction patterns known as symbiosis, parasitism, or predator-prey interactions. Pioneering advances in the design and study of synthetic multicellular systems have focused entirely on cross-talk within the same species. Quorum-sensing prokaryotic variants, engineered to produce lactones and broadcast this cell-density signal across a population, have been used to establish automated population control (1), programmed pattern formation (2, 3), or metabolic information processing in bacteria (4). Similar intrapopulation communication has also been successfully engineered in yeast using Arabidopsis thaliana-derived signaling pathways (5). Although these monospecies networks probe the design principles of their more complex natural counterparts, their extrapolation to mammalian cell communities or interspecies and interkingdom cross-talk remains limited. Furthermore, the use of nonvolatile small-molecule inducers as the broadcast signal in existing synthetic intercellular cross-talk systems necessitates that both sender and receiver cells reside in the same liquid environment. Sender cells transgenic for expression of alcohol dehydrogenase (ADH) produced acetaldehyde, ...

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“…Besides the Tet-MFP-, and ecdysone-regulated systems, different types of inducible lentiviral vectors have been generated based on the other chimeric regulatable systems, which include streptogramin-adjustable expression system derived from Streptomyces coelicolor (Mitta et al, 2004) and gaseous acetaldehyde-inducible expression system derived from Aspergillus nidulans (Hartenbach & Fussenegger, 2005). Although some endogenous cellular elements that respond to exogenous signals or stress have been adapted to HIV-1-and EIAV-derived lentiviral vectors (Beutelspacher et al, 2005;Hurttila et al, 2008;Parker et al, 2009), the pleiotropic effects exerted by the inducing agent would be a drawback in a human therapeutic context (Fussenegger, 2001).…”

Section: Other Regulatable Systemsmentioning

confidence: 99%