Vectors for gene expression in mammalian cells

Makrides, Savvas C.

doi:10.1016/s0167-7306(03)38002-0

“…Finally, the mammalian Shaker voltage-dependent K + channel was expressed in P. pastoris, which allowed efficient purification and high-resolution structure determination [63]. [73]. Alternatively, stable cell lines have been generated [74].…”

Section: Expression In Yeast Cellsmentioning

confidence: 99%

Structural Genomics and Drug Discovery

Anderson¹

2014

Methods in Molecular Biology

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Structure determination has already proven useful for lead optimization and direct drug design. The number of high-resolution structures available in public databases today exceeds 30,000 and will definitely aid in structure-based drug design. Structural genomics approaches covering whole genomes, topologically similar proteins or gene families are great assets for further progress in the development of new drugs. However, membrane proteins representing 70% of current drug targets are poorly characterized structurally. The problems have been related to difficulties in obtaining large amount of recombinant membrane proteins as well as their purification and structure determination. Structural genomics has proven successful in developing new methods in areas from expression to structure determination by studying a large number of target proteins in parallel.

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“…The promoter is of critical importance and should ideally be strong, constitutive, and active in a wide range of cell types. Many promoters have been used for gene expression in mammalian cells (Makrides, 2003), such as Rous sarcoma virus, SV40, EF1-a, and human ubiquitin, but the most commonly used remains the human cytomegalovirus (hCMV) promoter (Foecking and Hofstetter, 1986), which is one of the strongest eukaryotic promoters and is found in most commercial vectors.…”

Section: Plasmid Designmentioning

confidence: 99%

“…This can be obtained by fusing a signal sequence, usually at the 5¢ end of the cDNA sequence. Commonly used signal sequences come from erythropoietin, tissue plasminogen activator, albumin, or immunoglobulin (Makrides, 2003). Interestingly, in the case of the botulinum neurotoxins, there was no difference in the total antibody titers between the nonsecreted and the secreted forms of the same antigen, but the neutralizing titers showed much higher levels in the latter case after a single immunization, resulting in a much stronger protective effect versus a challenge (Trollet et al, 2009).…”

Section: Plasmid Designmentioning

confidence: 99%

Genetic Immunization with Plasmid DNA Mediated by Electrotransfer

Rochard

¹

,

Scherman

²

,

Bigey

³

2011

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The concept of DNA immunization was first advanced in the early 1990s, but was not developed because of an initial lack of efficiency. Recent technical advances in plasmid design and gene delivery techniques have allowed renewed interest in the idea. Particularly, a better understanding of genetic immunization has led to construction of optimized plasmids and the use of efficient molecular adjuvants. The field also took great advantage of new delivery techniques such as electrotransfer. This is a simple physical technique consisting of injecting plasmid DNA into a target tissue and applying an electric field, allowing up to a thousandfold more expression of the transgene than naked DNA. DNA immunization mediated by electrotransfer is now effective in a variety of preclinical models against infectious or acquired diseases such as cancer or autoimmune diseases, and is making its way through the clinics in several ongoing phase I human clinical trials. This review will briefly describe genetic immunization mediated by electrotransfer and the main fields of application.

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“…Recently, it was demonstrated that certain transmembrane proteins (RTP1, RTP2 and REEP1) [71], and the guanine nucleotide exchange factor Ric-8B [72] can facilitate the transport and enhance the expression of olfactory receptors. Transient expression has been conducted in various cell lines (BHK-21, CHO-K1, COS-7 and HEK293) [73]. Alternatively, stable cell lines have been generated [74].…”

Section: Transient and Stable Expressionmentioning

confidence: 99%

Structural genomics and drug discovery

Lundström¹

2007

J Cellular Molecular Medi

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Structure determination has already proven useful for lead optimization and direct drug design. The number of high-resolution structures available in public databases today exceeds 30,000 and will definitely aid in structure-based drug design. Structural genomics approaches covering whole genomes, topologically similar proteins or gene families are great assets for further progress in the development of new drugs. However, membrane proteins representing 70% of current drug targets are poorly characterized structurally. The problems have been related to difficulties in obtaining large amount of recombinant membrane proteins as well as their purification and structure determination. Structural genomics has proven successful in developing new methods in areas from expression to structure determination by studying a large number of target proteins in parallel.

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Vectors for gene expression in mammalian cells

Cited by 17 publications

References 66 publications

Structural Genomics and Drug Discovery

Structural Genomics and Drug Discovery

Genetic Immunization with Plasmid DNA Mediated by Electrotransfer

Structural genomics and drug discovery

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