Preface
Electroporation gene therapy, or gene electrotransfer, refers to the delivery of genetic
materials into target tissues or cells via electric pulses for the treatment or prevention of
disease. The genetic materials delivered by electroporation include oligoDNA, RNA, genes,
and chromosomes. This term is a by-product of electrochemotherapy, in which chemical
therapeutics or synthetic small molecules are delivered into targeted cells via electric pulses.
Simultaneous delivery of both genetic materials and chemical therapeutics is referred to as
electrochemogene therapy. The delivery of gentle/modest electric signal alone, without any
chemical therapeutic or genetic material, which is used to message tissue for functional
activation, is referred to as electrical (message) therapy, while the delivery of electric pulses
with high intensity/frequency without any other chemicals/genetic materials, which can be
used to remove bad tissue such as tumors, is referred to as irreversible electric therapy. The
focus of this book is to provide in-depth knowledge and hands-on protocols for the delivery
of naked DNA and small interfering RNA (siRNA) to the targeted cells, including fungus,
tissues, and animals, for the treatment of disease and biological studies. Therefore, this book
is primarily dedicated to electroporation gene transfer.
The first gene delivery via electroporation was carried out in 1982 by Professor Eberhard Neumann using an in vitro cell culture system. During the more than 30 years since
then, this technology has evolved greatly, thanks to the remarkable progress in genetic
sequencing, gene array analysis, gene cloning, gene synthesis, gene expression detection,
DNA manufacture, and discovery and synthesis of siRNA. This third edition of Electroporation Protocols: Microorganism, Mammalian System, and Nanodevice builds on the success of
the first two editions and on the progress made in the genetic delivery in single cells,
nanodevice, and microorganism.
Because of the rapid development in electroporation gene therapy, 100% of the chapters
in this third edition are completely new. Therefore, this book may be considered a continuation of the first two editions rather than a replacement in terms of the protocols. However,
the style and format used for the third edition are similar to those of the first two editions
and will be very familiar and accessible to investigators who have used the first two editions.
The major differences between this edition and the first two editions are the additions of
a large section on microorganism and a section nanodevice. The other major differences are
the substantial expansion of the section on applications in single cells because the electroporation of therapeutic genes has been used for preparing clinically used T cells for therapy.
This type of application is extremely valuable. These additions and expansions in the third
edition, unfortunately, required the sacrifice of chapters of the first two editions related to
large animal applications. For details of those topics, the first edition should serve very well.
This editor would like to acknowledge the coeditors of this book, Drs. Lingqian Chang
and Justin Teissie, who have done superb job in soliciting chapter authors, contributing
their works, and finalizing the contents. I also am grateful to all the authors who worked so
hard to get this book to press in a timely manner. Dr. Lingqian Chang also provided the art
for the cover. Finally, I am very grateful to my department chair, Richard Gorlick, MD, for
providing the time I needed to accomplish this work.
Houston, TX, USA
Shulin Li
v
Electroporation gene therapy, or gene electrotransfer, refers to the delivery of genetic
materials into target tissues or cells via electric pulses for the treatment or prevention of
disease. The genetic materials delivered by electroporation include oligoDNA, RNA, genes,
and chromosomes. This term is a by-product of electrochemotherapy, in which chemical
therapeutics or synthetic small molecules are delivered into targeted cells via electric pulses.
Simultaneous delivery of both genetic materials and chemical therapeutics is referred to as
electrochemogene therapy. The delivery of gentle/modest electric signal alone, without any
chemical therapeutic or genetic material, which is used to message tissue for functional
activation, is referred to as electrical (message) therapy, while the delivery of electric pulses
with high intensity/frequency without any other chemicals/genetic materials, which can be
used to remove bad tissue such as tumors, is referred to as irreversible electric therapy. The
focus of this book is to provide in-depth knowledge and hands-on protocols for the delivery
of naked DNA and small interfering RNA (siRNA) to the targeted cells, including fungus,
tissues, and animals, for the treatment of disease and biological studies. Therefore, this book
is primarily dedicated to electroporation gene transfer.
The first gene delivery via electroporation was carried out in 1982 by Professor Eberhard Neumann using an in vitro cell culture system. During the more than 30 years since
then, this technology has evolved greatly, thanks to the remarkable progress in genetic
sequencing, gene array analysis, gene cloning, gene synthesis, gene expression detection,
DNA manufacture, and discovery and synthesis of siRNA. This third edition of Electroporation Protocols: Microorganism, Mammalian System, and Nanodevice builds on the success of
the first two editions and on the progress made in the genetic delivery in single cells,
nanodevice, and microorganism.
Because of the rapid development in electroporation gene therapy, 100% of the chapters
in this third edition are completely new. Therefore, this book may be considered a continuation of the first two editions rather than a replacement in terms of the protocols. However,
the style and format used for the third edition are similar to those of the first two editions
and will be very familiar and accessible to investigators who have used the first two editions.
The major differences between this edition and the first two editions are the additions of
a large section on microorganism and a section nanodevice. The other major differences are
the substantial expansion of the section on applications in single cells because the electroporation of therapeutic genes has been used for preparing clinically used T cells for therapy.
This type of application is extremely valuable. These additions and expansions in the third
edition, unfortunately, required the sacrifice of chapters of the first two editions related to
large animal applications. For details of those topics, the first edition should serve very well.
This editor would like to acknowledge the coeditors of this book, Drs. Lingqian Chang
and Justin Teissie, who have done superb job in soliciting chapter authors, contributing
their works, and finalizing the contents. I also am grateful to all the authors who worked so
hard to get this book to press in a timely manner. Dr. Lingqian Chang also provided the art
for the cover. Finally, I am very grateful to my department chair, Richard Gorlick, MD, for
providing the time I needed to accomplish this work.
Houston, TX, USA
Shulin Li
v