Semliki Forest virus vectors for overexpression of 101 G protein-coupled receptors in mammalian host cells

Hassaı̈ne, Ghérici; Wagner, Renaud; Kempf, Juliette; Cherouati, N.; Hassaine, Nhouzha; Prual, Cécile; André, Nicolás; Reinhart, Christoph; Pattus, Franc; Lundström, Kenneth

doi:10.1016/j.pep.2005.06.007

Cited by 67 publications

(52 citation statements)

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“…In this context, especially SFV vectors have been used for the expression of topologically different proteins [14,19]. Particularly, the expression of membrane proteins has been very successful indicated by the expression of G protein-coupled receptors both at high levels [20] and in large numbers [21]. Moreover, they have demonstrated neuronspecific gene expression in primary neurons [22], in hippocampal slice cultures [23] and in vivo in rodent brain [24].…”

Section: Basics Of Alphavirusesmentioning

confidence: 99%

Alphavirus Vectors for Therapy of Neurological Disorders

Lundström¹

2012

J Stem Cell Res Ther

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Alphavirus vectors engineered for gene delivery and expression of heterologous proteins have been considered as valuable tools for research on neurological disorders. They possess a highly efficient susceptibility for neuronal cells and can provide extreme levels of heterologous gene expression. However, they generally generate short-term transient expression, which might limit their therapeutic use in many neurological disorders often requiring long-term even life-long presence of therapeutic agents. Recent development in gene silencing applying both RNA interference and microRNA approaches will certainly expand the application range. Moreover, alphaviruses provide interesting models for neurological diseases such as demyelinating and spinal motor diseases.

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Section: Basics Of Alphavirusesmentioning

confidence: 99%

Alphavirus Vectors for Therapy of Neurological Disorders

Lundström¹

2012

J Stem Cell Res Ther

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“…the most prominent target class is that of g-protein-coupled receptors (gpcrs), which cover about 60%-70% of recently developed drugs. 1 upon modulation by extracellular ligands, these cell surface receptors mediate the transduction of extracellular stimuli into intracellular signals. the hydrophobic core composed of seventransmembrane α-helices (7tm) is common to all gpcrs.…”

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Rapid Establishment of G-Protein-Coupled Receptor–Expressing Cell Lines by Site-Specific Integration

et al. 2011

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IntroductIon The conventional drug development process is expensive and time-consuming, and despite large investments in research and development activities, the success rate of bringing a new drug to the market is low. the most prominent target class is that of g-protein-coupled receptors (gpcrs), which cover about 60%-70% of recently developed drugs. 1 upon modulation by extracellular ligands, these cell surface receptors mediate the transduction of extracellular stimuli into intracellular signals. the hydrophobic core composed of seventransmembrane α-helices (7tm) is common to all gpcrs. 2 this large class comprises approximately 800 different receptors that are grouped in three families (classes a, B, and c).in general, the generation of cell lines stably expressing a transgene requires transduction of the host cell line and subsequent integration of the expression cassette into the host cell genome. this leads to a random distribution of integration sites.3 it is a fact that most genomic loci do not support high or consistent transgene expression. Both the level and stability of transgene expression are strongly dependent on the genomic surroundings of the integration sites. this "position effect" renders the generation of stable cell lines a cumbersome and often lengthy task involving extensive screening to identify those cell clones that display the desired properties. 4 the recombinase-mediated cassette exchange (rmce) strategy was introduced to overcome this limitation of cell line engineering (for review, see Wirth et al. 5 ). rmce can be best described as a two-step molecular "cut-and-paste" mechanism.6 in the first step, a reporter gene cassette is randomly integrated into a host cell line of interest. the expression of the reporter gene serves as a marker that permits screening for the desired expression characteristics (e.g., high and stable expression). once isolated and confirmed, those "tagged" clones function as master cell lines. a prerequisite for the use of such a master cell line is a single-copy integration of the tagging cassette. With the help of dna recombinases such as the cre, 7-9 the Flp recombinase, 10-12 or the phic31 integrase 13,14 the tagged genomic loci of the master cell lines can be recycled by the integration of any gene of interest. Because the recombinase-driven cassette exchange itself is a highly sitespecific event, all positive expression characteristics of the master cell line are transferred to the resulting subclonal producers. the aim of the study presented here is to employ the rmce strategy for the establishment of gpcr-expressing cell lines. the main application area of these cell lines is the drug 1 department of gene regulation and differentiation, hZi-helmholtz centre for infection research, Braunschweig, germany. 2 inscreeneX gmbh, Braunschweig, germany. 3 molecular devices (uK) ltd., Wokingham, uK. 4 european collections of cell cultures (ecacc), salisbury, Wiltshire, uK. the establishment of mammalian cell lines reliably expressing g-protein-coupled re...

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“…Forest Virus, que foi usado para a produção em larga escala de um receptor acoplado à proteína G, em células de mamífero cultivadas em suspensão em biorreator ou frasco de cultura tipo "spinner" (BLASEY et al, 2000;HASSAINE et al, 2006). Os métodos baseados em vírus, embora sejam mais eficientes, compreendem mais passos, sendo mais complicados e demandam mais tempo que os baseados em vetores plasmidiais.…”

Section: Um Dos Sistemas Virais Usados Amplamente Para Transfecção Trunclassified

Expressão da glicoproteína rábica utilizando pseudopartículas virais.

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Semliki Forest virus vectors for overexpression of 101 G protein-coupled receptors in mammalian host cells

Cited by 67 publications

References 45 publications

Alphavirus Vectors for Therapy of Neurological Disorders

Alphavirus Vectors for Therapy of Neurological Disorders

Rapid Establishment of G-Protein-Coupled Receptor–Expressing Cell Lines by Site-Specific Integration

Expressão da glicoproteína rábica utilizando pseudopartículas virais.

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