Table Of ContentMMeetthhooddss iinn MMoolleeccuullaarr BBiioollooggyy
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VOLUME 209
Transgenic
Transgenic
Mouse
Mouse
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Transgenic Mouse Methods and Protocols
M E T H O D S I N M O L E C U L A R B I O L O G YTM
John M. Walker, S E
ERIES DITOR
217. Neurogenetics: Methods and Protocols, edited by Nicholas 188. Epithelial Cell Culture Protocols, edited by Clare Wise, 2002
T. Potter, 2003 187. PCR Mutation Detection Protocols, edited by Bimal D. M.
216. PCR Detection of Microbial Pathogens: Methods and Pro- Theophilus and Ralph Rapley, 2002
tocols,edited by Konrad Sachse and Joachim Frey, 2003 186. Oxidative Stress Biomarkers and Antioxidant Protocols, ed-
215. Cytokines and Colony Stimulating Factors: Methods and ited by Donald Armstrong, 2002
Protocols,edited by Dieter Körholz and Wieland Kiess, 2003 185. Embryonic Stem Cells: Methods and Protocols, edited by
214. Superantigen Protocols, edited by Teresa Krakauer, 2003 Kursad Turksen, 2002
213. Capillary Electrophoresis of Carbohydrates, edited by 184. Biostatistical Methods, edited by Stephen W. Looney, 2002
Pierre Thibault and Susumu Honda, 2003 183. Green Fluorescent Protein: Applications and Protocols, edited
212. Single Nucleotide Polymorphisms: Methods and Protocols, byBarry W. Hicks, 2002
edited by Piu-Yan Kwok, 2003 182. In Vitro Mutagenesis Protocols, 2nd ed., edited by Jeff
211. Protein Sequencing Protocols, 2nd ed., edited by Bryan John Braman, 2002
Smith, 2003 181.Genomic Imprinting: Methods and Protocols, edited by
210. MHC Protocols, edited by Stephen H. Powis and Robert W. Andrew Ward, 2002
Vaughan, 2003 180. Transgenesis Techniques, 2nd ed.: Principles and Protocols,
209. Transgenic Mouse Methods and Protocols, edited by Marten edited by Alan R. Clarke, 2002
Hofker and Jan van Deursen, 2002 179. Gene Probes: Principles and Protocols, edited by Marilena
208. Peptide Nucleic Acids: Methods and Protocols, edited by Aquino de Muro and Ralph Rapley, 2002
Peter E. Nielsen, 2002 178. Antibody Phage Display: Methods and Protocols, edited by
207. Recombinant Antibodies for Cancer Therapy: Methods and Philippa M. O’Brien and Robert Aitken, 2001
Protocols.edited by Martin Welschof and Jürgen Krauss, 2002 177. Two-Hybrid Systems: Methods and Protocols, edited by Paul
206. Endothelin Protocols, edited by Janet J. Maguire and Anthony N. MacDonald, 2001
P. Davenport, 2002 176. Steroid Receptor Methods: Protocols and Assays, edited by
205. E. coli Gene Expression Protocols, edited by Peter E. Benjamin A. Lieberman, 2001
Vaillancourt, 2002 175. Genomics Protocols, edited by Michael P. Starkey and
204. Molecular Cytogenetics: Protocols and Applications, edited Ramnath Elaswarapu, 2001
byYao-Shan Fan, 2002 174. Epstein-Barr Virus Protocols, edited by Joanna B. Wilson and
203. In Situ Detection of DNA Damage: Methods and Protocols, Gerhard H. W. May, 2001
edited by Vladimir V. Didenko, 2002 173. Calcium-Binding Protein Protocols, Volume 2: Methods and
202. Thyroid Hormone Receptors: Methods and Protocols, edited Techniques,edited by Hans J. Vogel, 2001
by Aria Baniahmad, 2002 172. Calcium-Binding Protein Protocols, Volume 1: Reviews and
201. Combinatorial Library Methods and Protocols, edited by Case Histories, edited by Hans J. Vogel, 2001
Lisa B. English, 2002 171. Proteoglycan Protocols, edited by Renato V. Iozzo, 2001
200. DNA Methylation Protocols, edited by Ken I. Mills and Bernie 170. DNA Arrays: Methods and Protocols, edited by Jang B.
H, Ramsahoye, 2002 Rampal, 2001
199. Liposome Methods and Protocols, edited by Subhash C. Basu 169. Neurotrophin Protocols, edited by Robert A. Rush, 2001
and Manju Basu, 2002 168. Protein Structure, Stability, and Folding, edited by Kenneth
198. Neural Stem Cells: Methods and Protocols, edited by Tanja P. Murphy, 2001
Zigova, Juan R. Sanchez-Ramos, and Paul R. Sanberg, 2002 167. DNA Sequencing Protocols, Second Edition, edited by Colin
197. Mitochondrial DNA: Methods and Protocols, edited by William A. Graham and Alison J. M. Hill, 2001
C. Copeland, 2002 166. Immunotoxin Methods and Protocols, edited by Walter A. Hall, 2001
196. Oxidants and Antioxidants: Ultrastructural and Molecular 165. SV40 Protocols, edited by Leda Raptis, 2001
Biology Protocols, edited by Donald Armstrong, 2002 164. Kinesin Protocols, edited by Isabelle Vernos, 2001
195. Quantitative Trait Loci: Methods and Protocols, edited by 163. Capillary Electrophoresis of Nucleic Acids, Volume 2:
Nicola J. Camp and Angela Cox, 2002 Practical Applications of Capillary Electrophoresis, edited by
194. Posttranslational Modifications of Proteins: Tools for Functional Keith R. Mitchelson and Jing Cheng, 2001
Proteomics,edited by Christoph Kannicht, 2002 162. Capillary Electrophoresis of Nucleic Acids, Volume 1:
193. RT-PCR Protocols, edited by Joe O’Connell, 2002 Introduction to the Capillary Electrophoresis of Nucleic Acids,
192. PCR Cloning Protocols, 2nd ed., edited by Bing-Yuan Chen edited by Keith R. Mitchelson and Jing Cheng, 2001
and Harry W. Janes, 2002 161. Cytoskeleton Methods and Protocols, edited by Ray H. Gavin, 2001
191. Telomeres and Telomerase: Methods and Protocols, edited 160. Nuclease Methods and Protocols, edited by Catherine H.
byJohn A. Double and Michael J. Thompson, 2002 Schein, 2001
190. High Throughput Screening: Methods and Protocols, edited 159. Amino Acid Analysis Protocols, edited by Catherine Cooper,
byWilliam P. Janzen, 2002 Nicole Packer, and Keith Williams, 2001
189. GTPase Protocols: The RAS Superfamily, edited by Edward 158. Gene Knockoout Protocols, edited by Martin J. Tymms and
J. Manser and Thomas Leung, 2002 Ismail Kola, 2001
M E T H O D S I N M O L E C U L A R B I O L O G YTM
Transgenic Mouse
Methods and Protocols
Edited by
Marten H. Hofker
Department of Molecular Genetics
University of Maastricht
Maastricht, The Netherlands
Jan van Deursen
Department of Pediatrics and Adolescent Medicine
Mayo Clinic, Rochester, MN
Humana Press Totowa, New Jersey
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Transgenic mouse methods and protocols / edited by Marten H. Hofker, Jan van Deursen.
p. cm. -- (Methods in molecular biology ; 209)
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ISBN 0-89603-915-3 (alk. paper)
1. Transgenic mice--Laboratory manuals. I. Hofker, Marten H. II. Deursen, Jan van, 1930– III. Methods
in molecular biology (Totowa, N.J.) ; v. 209
QH442.6 .T724 2002
616'.93--dc21
2002068581
Preface
Mice play an important role in biomedical research. Genetically, the mouse
is very similar to the human species, and most human genes are conserved in the
mouse. Moreover, the arrangement of genes on chromosomes is similar in humans
and mice. Mouse models have been established for a wide variety of human
disorders including cancer, cardiovascular diseases, and behavioral disorders.
Transgenic Mouse Methods and Protocols covers the production and analysis of
transgenic and knockout mice. The book also includes a review of mouse models
for a wide range of genetic aberrations associated with human disease.
Many researchers played a crucial role in developing mouse technology to
the excellent state of art that has now been achieved. Landmarks include the
generation of (i) transgenic mice, (ii) pluripotent embryonic stem (ES) cell cultures,
(iii) gene knockout mice, (iv) tissue-specific knockouts, and (v) systems for
inducible gene expression mice. Most of these landmarks have not been achieved
in other mammalian systems with a comparable efficiency. In part, this is attributable
to the availability of hundreds of different inbred mouse strains, which allowed
researchers to choose from a wide range of strains while establishing these
technologies.
Transgenic Mouse Methods and Protocols has essentially the same format
as previous volumes of the series Methods in Molecular Biology. Since mouse
technology offers a wide range of possibilities, most chapters will provide the
rationale for choosing the given protocol, which is then described in step-by-step
detail. In addition, two chapters will describe alternative strategies for studying
gene function in the mouse, i.e., random mutagenesis and viral gene transduction.
The book can be roughly divided into three topics: a general introduction describing
how to deal with mice and how to generate transgenics; a technology part where
new methods will be presented, including conditional and induced knockout and
transgenic mice; and the final chapters, which highlight working with mice in
several major research areas, including cancer, immunology, and atherosclerosis.
We would like to thank the authors for their excellent contributions and Wilma
Claessen for editorial assistance. We are very grateful to Be Wieringa, Anton Berns,
and Robin Lovell Badge for leading us into the world of gene-targeting and ES
cell technology.
Marten Hofker
Jan van Deursen
v
Contents
Preface .............................................................................................................v
Contributors .....................................................................................................ix
Color Plates ................................................................................................... xiii
1 Introduction: The Use of Transgenic Mice in Biomedical Research
Marten H. Hofker....................................................................................1
2 Genetic Modification of the Mouse:
General Technology; Pronuclear and Blastocyst Injection
J. Willem Voncken.................................................................................9
3 In Vitro Fertilization/Cryopreservation
ˆ
Susan Marschall 0and Martin Hrabe de Angelis.............................35
4 Transgene Design
J. Willem Voncken...............................................................................51
5 Generating Conditional Mouse Mutants
via Tetracycline-Controlled Gene Expression
Kai Schönig and Hermann Bujard.....................................................69
6 Manipulating Large Insert Clones for Transgenesis
Sally P. A. McCormick, Catherine Y. Y. Liu, Stephen G. Young,
and Lars B. Nielsen.......................................................................105
7 Reporter Molecules in Genetically Engineered Mice
Thomas L. Saunders.........................................................................125
8 Gene Targeting in Mouse Embryonic Stem Cells
Jan van Deursen................................................................................145
9 Conditional Knockout Mice
Ralf Kühn and Frieder Schwenk......................................................159
10 Knockin Approaches
Anton J. M. Roebroek, Xiaosheng Wu, and Richard J. Bram......187
11 Aggregation of Embryos and Embryonic Stem Cells
Ronald Maatman, Marina Gertsenstein, Emile de Meijer,
Andras Nagy, and Kristina Vintersten........................................201
12 Andenovirus-Mediated Gene Transfer
Ko Willems van Dijk, Kyriakos E. Kypreos,
Christine d’Oliveira, and Frits J. Fallaux....................................231
vii
viii Contents
13 Random ENU Mutagenesis
Dian Soewarto, Véronique Blanquet,
ˆ
and Martin Hrabe de Angelis.......................................................249
14 Autopsy and Histologic Analysis of the Transgenic Mouse
Marion J. J. Gijbels and Menno P. J. de Winther..........................267
15 Bone Marrow Transplantations to Study Gene Function
in Hematopoietic Cells
Menno P. J. de Winther and Peter Heeringa..................................281
16 Quantification of Atherosclerosis in Mice
Alan Daugherty and Stewart C. Whitman.......................................293
17 Genetically Modified Mice in Cancer Research
David A. Largaespada.......................................................................311
18 Neuropathobiology in Transgenic Mice:
The Case of Alzheimer’s Disease
Jo Van Dorpe, Kurt Spittaels, Chris Van den Haute,
Ilse Dewachter, Dieder Moechars, Hugo Geerts,
and Fred Van Leuven....................................................................333
Index ............................................................................................................363
Contributors
V B • GSF Research Centre for Environment and Health,
ÉRONIQUE LANQUET
Institute of Experimental Genetics, Neuherberg, Germany
R J. B • Department of Pediatrics and Adolescent Medicine, Mayo
ICHARD RAM
Clinic, Rochester, MN
H B • Zentrum für Molekulare Biologie der Universität
ERMANN UJARD
Heidelberg (ZMBH), Heidelberg, Germany
A D • Gill Heart Institute, Division of Cardiovascular
LAN AUGHERTY
Medicine, University of Kentucky, Lexington KY
J D • Department of Pediatrics and Adolescent Medicine,
AN VAN EURSEN
Mayo Clinic, Rochester, MN
I D • Experimental Genetics Group, Center for Human
LSE EWACHTER
Genetics, Flemish Institute for Biotechnology, Katholieke Universiteit
Leuven, Leuven, Belgium
K W D • Departments of General Internal Medicine and of
O ILLEMS VAN IJK
Human Genetics, Leiden University Medical Center, Leiden, The
Netherlands
J V D • Experimental Genetics Group, Center for Human
O AN ORPE
Genetics, Flemish Institute for Biotechnology, Katholieke Universiteit
Leuven, Leuven, Belgium
F J. F • Department of Human Genetics, Leiden University
RITS ALLAUX
Medical Center, Leiden, The Netherlands
H G • Janssen Research Foundation, Beerse, Belgium
UGO EERTS
M G • Samuel Lunenfeld Research Institute, Mount Sinai
ARINA ERTSENSTEIN
Hospital, Toronto, Canada
M J. J. G • Department of Molecular Genetics, Universiteit
ARION IJBELS
Maastricht, Maastricht, The Netherlands
C V H • Experimental Genetics Group, Center for Human
HRIS AN DEN AUTE
Genetics, Flemish Institute for Biotechnology, Katholieke Universiteit
Leuven, Leuven, Belgium
P H • Department of Clinical and Experimental Immunology,
ETER EERINGA
Universiteit Maastricht, Maastricht, The Netherlands
M H. H • Department of Molecular Genetics, Universiteit
ARTEN OFKER
Maastricht, Maastricht, The Netherlands
ˆ
M H A • GSF Research Centre for Environment and
ARTIN RABE DE NGELIS
Health, Institute of Experimental Genetics, Neuherberg, Germany
R K • Artemis Pharmaceuticals GmbH, Cologne, Germany
ALF ÜHN
ix
