Gene Transfer And Expression In Mammalian Cells

This volume provides a broad, state-of-the-art coverage of diverse technical topics in gene expression in mammalian cells, including the development of vectors for production of proteins in cultured cells, in transgenic animals, vaccination, and gene therapy; progress in methods for the transfer of genes into mammalian cells and the optimization and monitoring of gene expression; advances in our understanding and manipulation of cellular biochemical pathways that have a quantitative and qualitative impact on mammalian gene expression; and the large-scale production and purification of proteins from cultured cells.

Experienced researchers describe in step-by-step detail methods that have proven most useful in delivering genes to mammalian cells. Volume 1 focuses on gene delivery by a variety of chemical and physical methods, including ultrasound, biolistics, peptides, PNA clamps, liposomes, microinjection, electroporation, particle bombardment, dendrimers, and hydrodynamics. Volume 2 details procedures for delivering genes to cells in vitro and in vivo, including the use of lentiviral vectors.

Biology is the study of living things. The classical approach might be described as holistic and descriptive, whereas the modern molecular - proach aims to be investigative, reductionist, and mechanistic . Genes contain all the information for the structure of all living things ; thus, the understanding of how genes are regulated is an important step toward understanding the nature of living things. The study of gene regulation has been made more tractable by the design of simple expe- mental models in which a single gene can be isolated from the milieu of the organism. The new science of molecular biology has introduced techniques that permit the design of such experimental models. In - sence, the genome of the organism is dissected in such a manner that specific genes may now be introduced into an appropriate cell line . Subsequent analysis of the proteins expressed from the genes under study results in the identification of the regulatory DNA sequences .

A practical manual of protocols for achieving expression of foreign genes in mammalian cells. It includes some very new techniques such as PCR-based expression. The author gives a theoretical introduction to the protocols and compares the strengths and weaknesses.

Gene transfer to animal cells was first achieved more than thirty years ago. Since then, transformation technology has developed rapidly, resulting in a multitude of techniques for cell transformation and the creation of transgenic animals. As with any expanding technology, it becomes difficult to keep track of all the developments and to find a concise and comprehensive source of information that explains all the underlying principles. Gene Transfer to Animals Cells addresses this problem by describing the principles behind gene transfer technologies, how gene expression is controlled in animal cells and how advanced strategies can be used to add, exchange or delete sequences from animal genomes in a conditional manner. A final chapter provides an overview of all the applications of animal cell transformation in farming, medicine and research.

Genetic analysis of microbial systems provided us with the foundation for un derstanding gene structure, expression, and regulation. It was long felt that the ability to generate mutants and conduct genetic studies in mammalian systems would prove to be equally useful. However, genetic analysis based on sexual systems is difficult in mammals because of the long generation times and the inability to perform controlled matings. As a result, genetic analysis of mam malian systems had to await the development of parasexual systems. This book is an attempt to bring together descriptions of a number of these parasexual systems. A common theme of all the parasexual systems is the transfer of genetic information from a defined source into a specific cell type. This volume deals with a number of methods of gene transfer into mammalian cells. The early methods of gene transfer involved transfer of relatively large amounts of genetic information. These include somatic cell hybridization, microcell fusion, and chromosome transfer, which constitute the first part of this book. Each of these methods has already proven to be of enormous value in arriving at a genetic understanding of the mammalian genome. Development of recombinant DNA methods, and the ability to introduce purified DNA into mammalian cells, has had a significant impact on our ability to dissect important aspects of mammalian gene expression and regulation. The second part of this book deals with gene transfer systems involving defined nucleic acid sequences.

During the first half century of genetics, coinciding with the first half of this cen tury, geneticists dreamt of the repair of genetic disease by altering or replacing defective genes. H. J. Muller wrote of the great advantages of mutations, "nanoneedles" in his apt term, for delicately probing physiological and chemical processes. In the same spirit, genes could be used to provide treatments of needle point delicacy. Yet, during this period no realistic possibility appeared; it remained but a dream. The situation changed abruptly at the half century. Microbial genetics and its offshoot, cell culture genetics, provided the route. Pneumococcus transformation showed that exogenous DNA could become a permanent part of the genome; yet attempts to reproduce this in animals produced a few tantalizing hints of success, but mostly failures. Transduction, using a virus as mediator, offered a better op portunity. The fITSt reproducible in vivo gene therapy in a whole animal came in 1981. This was in Drosophila, with a transposable element as carrier. Flies were "cured" of a mutant eye color by incorporation of the normal allele, and the effect was transmissible, foreshadowing not only somatic, but germ line gene therapy. At the same time, retroviruses carrying human genes were found to be ex tremely efficient in transferring their contents to the chromosomes of cultured cells.