遗传学实验技术

Adenoviral vector (AdV) of the third generation also known as helper-dependent adenoviral vector (HDV) is an attractive delivery system for gene therapy applications. However, obtaining high quality-grade HDV in sufficient amount remains a challenge that hampers the extensive use ...

The most efficient system to introduce genes of interest within the adenovirus genome is by homologous recombination in microorganisms. In this chapter, the most popular procedures are described: two for homologous recombination in Escherichia coli, and one in yeast. Main differenc ...

As a result of the growing interest in the use of viruses for gene therapy and vaccines, many virus-based products are being developed. The manufacturing of viruses poses new challenges for process developers and regulating authorities that need to be addressed to ensure quality, efficacy, a ...

Gene therapy based on the use of viral vectors is entirely dependent on the use of animal cell lines, mainly of mammalian origin, but also of insect origin. As for any biotechnology product for clinical use, viral �vectors have to be produced with cells derived from an extensively characterized cell b ...

Despite three decades of huge progress in molecular genetics, in cloning of disease causative gene as well as technology breakthroughs in viral biotechnology, out of thousands of gene therapy clinical trials that have been initiated, only very few are now reaching regulatory approval. We s ...

Viral vector is the most effective means of gene transfer to modify specific cell type or tissue and can be manipulated to express therapeutic genes. Several virus types are currently being investigated for use to deliver genes to cells to provide either transient or permanent transgene expr ...

Advances in gene therapy are increasingly leading to clinical assessment in many fields of medicine with diverse approaches. The basic science stems from approaches aimed at different functions such as correcting a missing/abnormal gene, altering the proportion or expression of no ...

The twofold distinction between interventions into the germ line and interventions into somatic cells on the one hand and between the treatment of diseases and enhancement on the other hand resulted in the concept of somatic gene therapy. There is a nearly unanimous agreement that somatic ge ...

Gene transfer of specific growth factors is suitable for inducing chondrogenic differentiation of �mesenchymal cells to be used for cartilage regeneration. However, extent and quality of repair tissue formation also depend on biomechanical and metabolic influences that can only ...

In vivo targeted gene delivery to hematopoietic stem cells (HSCs) would mean a big step forward in the field of gene therapy. This would imply that the risk of cell differentiation and loss of homing/�engraftment is reduced, as there is no need for purification of the target cell. In vivo gene delivery also ...

Measles viruses have shown potent oncolytic activity as a therapeutic against a variety of human cancers in animal models and are currently being tested in clinical trials in patients. In contrast to using measles virus as a vaccine, oncolytic activity depends on high concentrations of infe ...

Herpes simplex virus type 1 (HSV-1) is a human pathogen whose lifestyle is based on a long-term dual interaction with the infected host, being able to establish both lytic and latent infections. The virus genome is a 153 kbp double-stranded DNA molecule encoding more than 80 genes. The interest of HSV-1 as ...

Baculovirus expression vector system (BEVS) is well known as a feasible and safe technology to produce recombinant (re-)proteins in a eukaryotic milieu of insect cells. However, its proven power in gene delivery and gene therapy is still poorly recognized. The basis of BEVS lies in large envelo ...

Adeno-associated virus (AAV)-based vectors expressing therapeutic gene products have shown great promise for human gene therapy. A major challenge for translation of promising research to clinical development is the establishment of appropriate quality control (QC) test met ...

Adeno-associated virus (AAV) is one of the most promising vectors for gene therapy. There are several ways of producing AAV vectors but large-scale production of this vector remains a major challenge. Virovek developed a novel method of expressing the AAV Rep and Cap genes in insect cells mediat ...

The human genome project has predicted that the human genome encodes about 35,000 genes (1,2). These genes are not uniformly expressed in all of the cells. Some of them are expressed in most of the cells, but others are cell-or tissue-specific. It has been estimated that about 10,000 genes are expressed in a c ...

Synthesis of complementary DNA (cDNA) by reverse transcription (RT) is a key step in investigating specific gene expression of a single transcript by RT-PCR (polymerase chain reaction) or to study the more complex profiles of gene expression in a biological sample using cDNA library or other t ...

Since the first report on complementary DNA (cDNA) cloning in 1972 (1), the technology has been developed into a powerful and universal tool in the isolation, characterization, and analysis of both eukaryotic and prokaryotic genes. However, the conventional methods of cDNA cloning requi ...

In Chapter 10, we describe two methods of complementary DNA (cDNA) synthesis and amplification. In this chapter, we will refer to these two methods of cDNA synthesis and amplification as “method A” (the technique based on classical double-stranded cDNA synthesis and subsequent adapter lig ...

In the past, the construction of complementary DNA (cDNA) libraries has served as a key technology for the discovery of biologically interesting genes (1). For mammalian genes, this is true even in the postgenome sequencing era. The prediction of protein-coding sequences solely from the gen ...