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DNA Transfer to Cultured Cells
Cell Biology International, 2000, Vol. 24, No. 7, 493 doi:10.1006/cbir.1999.0540, available online at http://www.idealibrary.com on
BOOK REVIEW DNA Transfer to Cultured Cells Edited by RAVID & FRESHNEY Wiley, U.K., 1998. Pp. 312. £45.50. ISBN 0-47116-572-7 Introduction of nucleic acids into cells has profoundly improved the way in which genes are studied within eukariotic cells and tissues. Today, the study of genetic regulation, protein expression and function in mammalian cells, the generation of transgenic organisms and gene therapy are enabled thanks to the growing advancements in gene transfer technologies. The different ways of introducing genes into cultured cells and tissues can be broadly classified as either chemical reagents or physical methods. The materials discussed in the book are centred on methods of DNA transfer utilizing chemical reagents. The first two chapters are devoted to virus infection, which is today one of the most efficient ways to transfer genes into mammalian cells in culture and tissues. Four other chapters focus on the calcium phosphate precipitation, which is one of today’s most widely used transfection methods in cultured mammalian cells. The book also includes three interesting articles on the DEAE-Dextran and liposomal vehicles. These methods are based on the ability of the cationic polymer and lipids to associate with negatively charged nucleic acid, and the artificial liposome to undergo endocytosis. It is unclear, however, how the nucleic acids are released from the endosomes and traverse the nucleic membrane. Two chapters extensively describe the electroporation transfection technique, which is often used for cell types that are particularly recalcitrant to milder methods of gene transfer. It would also have been useful to include some headings dedicated to microinjection or biolistic particle delivery into cultured cells or tissues. Beside these different methods of introducing genes into cells, the book provides four sections describing the use of the baculovirus expression system as a popular route to the production of high levels of eukaryotic proteins, zebrafish embryos for studies of vertebrate development and toxicology, mammalian cells transfected with yeast chromo1065–6995/00/070493+01 $35.00/0
somes and gene amplification of DHFR and coamplification of co-transfected DNA when cells are exposed to methotrexate. The book provides a very comprehensive overview of the most commonly used techniques of gene transfer today, grouped into 14 chapters written by 30 contributors whose expertise spans the field of cellular and molecular biology. Interestingly, each chapter also provides considerably detailed protocols for preparation of DNA, primary tissue culture and gene transfer procedure. It is worth noting that special sections at the end of each chapter, as well as at the end of the book itself, list suppliers for necessary reagents and materials. In a few words, this is an easy-to-use guide for the practice of gene transfer into cultured cells that every scientist may want to find on his bench. Due to the complexity and rapid growth of technology research on DNA transfer to cultured cells, only few texts have attempted to give an overview of this field. One should therefore be grateful to K. Ravid and R. I. Freshney for their well-written and comprehensive book. I must, however, confess some minor disappointments. It would have been interesting if some complimentary approaches had been included. First, I would have liked to start reading this book with an introductory chapter that would have presented an historical overview and latest developments of the transfection techniques. Also, most of the preparations used in this book are immortalized cell lines and we know that one of the most important challenges today is to transfer DNA into more recalcitrant cells and tissues such as primary or organotypic neuronal cultures. Also, very recent and efficient physical methods of gene transfer are not presently approached. In conclusion, this is a very useful book that may enable every one of us to improve transfection efficiency by following the recommendations and protocols that are displayed in a very comprehensive way in this book. It may, however, be essential to optimize each specific transfection condition to gain optimal efficiencies and the important parameters to optimize are clearly presented for most of the techniques approached by the different contributors. Laurent Fagni Montpellier December 1999 2000 Academic Press
BOOK REVIEW DNA Transfer to Cultured Cells Edited by RAVID & FRESHNEY Wiley, U.K., 1998. Pp. 312. £45.50. ISBN 0-47116-572-7 Introduction of nucleic acids into cells has profoundly improved the way in which genes are studied within eukariotic cells and tissues. Today, the study of genetic regulation, protein expression and function in mammalian cells, the generation of transgenic organisms and gene therapy are enabled thanks to the growing advancements in gene transfer technologies. The different ways of introducing genes into cultured cells and tissues can be broadly classified as either chemical reagents or physical methods. The materials discussed in the book are centred on methods of DNA transfer utilizing chemical reagents. The first two chapters are devoted to virus infection, which is today one of the most efficient ways to transfer genes into mammalian cells in culture and tissues. Four other chapters focus on the calcium phosphate precipitation, which is one of today’s most widely used transfection methods in cultured mammalian cells. The book also includes three interesting articles on the DEAE-Dextran and liposomal vehicles. These methods are based on the ability of the cationic polymer and lipids to associate with negatively charged nucleic acid, and the artificial liposome to undergo endocytosis. It is unclear, however, how the nucleic acids are released from the endosomes and traverse the nucleic membrane. Two chapters extensively describe the electroporation transfection technique, which is often used for cell types that are particularly recalcitrant to milder methods of gene transfer. It would also have been useful to include some headings dedicated to microinjection or biolistic particle delivery into cultured cells or tissues. Beside these different methods of introducing genes into cells, the book provides four sections describing the use of the baculovirus expression system as a popular route to the production of high levels of eukaryotic proteins, zebrafish embryos for studies of vertebrate development and toxicology, mammalian cells transfected with yeast chromo1065–6995/00/070493+01 $35.00/0
somes and gene amplification of DHFR and coamplification of co-transfected DNA when cells are exposed to methotrexate. The book provides a very comprehensive overview of the most commonly used techniques of gene transfer today, grouped into 14 chapters written by 30 contributors whose expertise spans the field of cellular and molecular biology. Interestingly, each chapter also provides considerably detailed protocols for preparation of DNA, primary tissue culture and gene transfer procedure. It is worth noting that special sections at the end of each chapter, as well as at the end of the book itself, list suppliers for necessary reagents and materials. In a few words, this is an easy-to-use guide for the practice of gene transfer into cultured cells that every scientist may want to find on his bench. Due to the complexity and rapid growth of technology research on DNA transfer to cultured cells, only few texts have attempted to give an overview of this field. One should therefore be grateful to K. Ravid and R. I. Freshney for their well-written and comprehensive book. I must, however, confess some minor disappointments. It would have been interesting if some complimentary approaches had been included. First, I would have liked to start reading this book with an introductory chapter that would have presented an historical overview and latest developments of the transfection techniques. Also, most of the preparations used in this book are immortalized cell lines and we know that one of the most important challenges today is to transfer DNA into more recalcitrant cells and tissues such as primary or organotypic neuronal cultures. Also, very recent and efficient physical methods of gene transfer are not presently approached. In conclusion, this is a very useful book that may enable every one of us to improve transfection efficiency by following the recommendations and protocols that are displayed in a very comprehensive way in this book. It may, however, be essential to optimize each specific transfection condition to gain optimal efficiencies and the important parameters to optimize are clearly presented for most of the techniques approached by the different contributors. Laurent Fagni Montpellier December 1999 2000 Academic Press