Mutagenesis of the Mouse Genome

98 Pages·2004·1.297 MB·English

by  Monica J. Justice (auth.), Monica J. Justice, Mary Bedell (eds.)

MUTAGENESIS OF THE MOUSE GENOME
Georgia Genetics Review II
VOLUME 2
Mutagenesis of the Mouse Genome
Edited by
MONICA JUSTICE AND MARY BEDELL
Reprinted from Genetica, Volume 122(1), 2004
KLUWER ACADEMIC PUBLISHERS
DORDRECHT =BOSTON =LONDON
CONTENTS
Foreword 1
Monica Justice
From the Atomic Age to The Genome Project 3
M.J. Justice
Chemical mutagenesis of the mouse genome: an overview 9
J.-L. Gue´net
Effectsofmalegerm-cellstageonthefrequency,nature,andspectrumofinducedspecific-locus 25
mutationsinthemouse
L.B. Russell
Allelic mutations of the sodium channel SCN8A reveal multiple cellular and physiological 37
functions
M.H. Meisler, N.W. Plummer, D.L. Burgess, D.A. Buchner, L.K. Sprunger
Towardsamutantmapofthemouse–newmodelsofneurological,behavioural,deafness,bone, 47
renal and blood disorders
S. Rastan, T. Hough, A. Kierman, R. Hardisty, A. Erven, I.C. Gray, S. Voeling, A. Isaacs,
H.Tsai,M.Strivens,R.Washbourne,C.Thornton,S.Greenaway,M.Hewitt,S.McCormick,
R. Selley, C. Wells, Z. Tymowska-Lalanne, P. Roby, P. Mburu, D. Rogers, J. Hagan,
C.Reavill,K.Davies,P.Glenister,E.M.C.Fisher,J.Martin,L.Vizor,M.Bouzyk,D.Kelsell,
J.-L.Guenet,K.P.Steel,S.Sheardown,N.Spurr,I.Gray,J.Peters,P.M.Nolan,A.J.Hunter,
S.D.M.Brown
Implementing large-scale ENU mutagenesis screens in North America 51
A.T. Clark, D. Goldowitz, J.S. Takahashi, M.H. Vitaterna, S.M. Siepka, L.L. Peters,
W.N. Frankel, G.A. Carlson, J. Rossant, J.H. Nadeau, M.J. Justice
Genetic mapping and ENU mutagenesis 65
D.R. Beier, B.J. Herron
The Mouse Phenome Project 71
M.A. Bogue, S.C. Grubb
Genetically altered mice: phenotypes, no phenotypes, and Faux phenotypes 75
S.W. Barthold
Visualizing the laboratory mouse: capturing phenotype information 89
M. Strivens, J.T. Eppig
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ISBN 1-4020-2875-X
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1
Foreword
TheSecondGeorgiaGeneticsSymposiumwasheld color.Soonafter,hejoinedthestaffofTheJackson
in September 2000, and the development of this Laboratory in Bar Harbor,Maine.
booktookplaceoverthenearly4 yearsthatensued. Much of Bill’s research at the lab was centered
During this time, many advances in the Genome around investigating phenotypic variability within
Project and mouse mutagenesis were made. In the highly inbred strains, and in that connection he
book overview, we discuss the development of the developed the technique of ovarian transplanta-
GenomeProject(which isthecontextforthesym- tion (even using embryonic donors) and a genetic
posium), the role the mouse was playing at that scheme whereby graft compatibility could be
time, how that role has evolved, and how the combined with the ability to distinguish offspring
chaptersofthebookaddressissuesinmousefunc- from donor and regenerated host ovaries. His
tional genetics. Many of the chapters in this book work was influenced by the second World War,
willprovideusefulresourcesforyearstocome. first because The Jackson Laboratory turned into
Of greater impact, our keynote speaker, the a production colony for the military, primarily to
mutagenesis pioneer William L. (Bill) Russell, produce mice for typhoid testing, and secondly,
passed away on July 23, 2003. I am one of an becausehebecameconsultanttoanoff-siteproject
exclusivegroupofmousegeneticistswhocanboast on testing the effects of radiation on mice. Bill
of having had the opportunity to work at Oak sharedmemoriesofhowscientistswereawarethat
Ridge National Laboratory, and to interact with theworldwas changing just before WorldWarII,
Bill and his wife Liane Brauch (Lee) Russell. Pro- partially because all articles that mentioned the
lific and unique authors in the field of mouse word ‘uranium’ had disappeared from Science
genetics and mutagenesis, their papers reflect a magazine. Bill was attending the International
density of genetic knowledge that is too often Genetics Congress in Edinburgh when many sci-
overlooked in today’s fast-paced molecular world. entistsdepartedthemeetingmid-courseduetothe
Bill was a resource of scientific and historical advent of the war, which started the day after the
information, whose memory was extraordinary. meeting ended, September 1, 1939. The advent of
Becausehelikedthequietenvironmentofhishome the atomic age during World War II had great
in Oak Ridge, he would often invite me over for significance for Bill’s research career.
lunch (always cream of mushroom soup, home- Together, Bill and Lee loved hiking and
grown tomato and cheese sandwiches made with canoeing in the Tennessee wilderness, and helped
homemadebread)anddiscussionsofscience,music to establish the Tennessee Citizens for Wilderness
and history. He was born in 1910, before the start Planning (TCWP). Bill and Lee were leaders in
ofWorldWarI,andgrewupinNewhaven,onthe saving two nearby river gorges from dams and
south coast of England. His memories of the first securing their protection by having them added to
World War were so descriptive that I could hear the National Park System. They also developed
thedistantsoundofthebombsacrossthechannel, the North Ridge Trail, which protected the
andseethestealthyfloatingzeppelinsontheirway northern greenbelt of Oak Ridge, while providing
to drop bombs on London. Later, he attended a beautiful well-maintained path for hiking that I
Oxford University and then won a foreign schol- enjoyed when I lived in Oak Ridge. Bill loved
arship to Amherst College in America, where he music, photography and woodworking, in addi-
pursued his interest in genetics by working on a tion to the wilderness. Bill’s devotion to his wife
small Drosophila research project (on Notch!) in Lee was obvious for all to see. They shared not
thelabofProfessorHaroldPlough.Hethenmoved onlymanycommoninterests,amongthemscience
to the University of Chicago to pursue a doctoral and the environment, but a special bond that few
degreein thelaboratoryof thefamous population othershave.Wededicatethisbooktohismemory.
geneticistSewallWright,earninghisdegreein1937
on the physiological genetics of guinea pig coat Monica Justice
Genetica 122: 3–7,2004. 3
(cid:1)2004KluwerAcademicPublishers.PrintedintheNetherlands.
From the Atomic Age to The Genome Project
Monica J. Justice
DepartmentofMolecularandHumanGenetics,BaylorCollegeofMedicine,OneBaylorPlazaS413,Houston,
TX 77030, USA (Phone: [phone]; Fax: [phone]; E-mail:[email protected])
Key words: functional genomics, Human Genome Project, mouse genome sequence, mouse mutagenesis,
specific locus test
Introduction come one of the most powerful of model organ-
isms to help to annotate the functions of genes in
The Human Genome Project has changed our mammals.
approach to biology. Sequence data are being ac-
quired from multiple organisms at a phenomenal
pace.The sequencing hasnotonlydisclosedmany
novel genes, but allows for the evolutionary com- The Genome Project
parisons of genes and their expression. The future
challenge will be to discover how their regulation In the U.S., interest in sequencing the human
and functions have changed over evolutionary genome began as early as 1984, and two meetings
distance and speciation. to assess feasibility were held in 1985. Pilot pro-
Mutagenesisisbeingcarriedoutinmanymodel jects were funded within The Department of En-
organismstodiscovernewgenefunctions,because ergy(DOE)nationallaboratoriesin1986.In1987,
geneticvariantshaveenabledtheunderstandingof a congressionally chartered DOE advisory com-
gene regulation, and biochemical and develop- mittee recommended a 15-year multidisciplinary
mental pathways. The mouse is the first mamma- undertaking to map and sequence the human
lian organism to be subjected to large-scale genome. In response to this recommendation, the
mutagenesis, and will permit key discoveries be- DOE designated several human genome centers,
cause of its many morphological, physiological, and the NIH National Institute of General Medi-
developmental and reproductive similarities to the cal Science (NIGMS) began funding genome pro-
human. A comparative analysis of gene functions jects.Theprojectdidnotofficiallybegin,however,
thatincludesthelaboratorymousewillexpandour until 1990, when the DOE and NIH presented a
knowledge of the genetic, developmental and joint 5 year Human Genome Project plan to
physiologicaldifferencesbetweenamammalanda Congress.A‘workingdraft’oftheDNAsequence
fruit fly. of the human genome was completed in
The Second Annual Georgia Genetics Sympo- 2000, concurrent with the Georgia Genetics Sym-
sium: Mutagenesis of the Mouse Genome focused posium.
on new directions in functional genetics in the In anticipation of the utility of the mouse se-
mouse. Our goal in this meeting was to invite quence to annotate the human genome sequence,
speakers who were tackling the genome using a the mouse community prepared several initiatives
variety of mutagenesis techniques. Further, the to plan for the ‘post-genome’ era. In 1995, at the
development of resources and infrastructure for 9th Annual Mouse Genome Conference, the an-
managing mutant mouse strains was needed. nual meeting of the International Mammalian
Therefore, we decided to build a program that GenomeSociety,agroupofmousegeneticistsfrom
focused on the generation, mapping and manage- five countries met in a small room to discuss the
ment of mouse mutant resources. In doing so, we power of the chemical mutagen, N-ethyl-N-nitro-
hoped to demonstrate that the mouse would be- sourea(ENU)tomakepointmutationsthroughout
4
the mouse genome. In 1998, the NIH convened a chapter for this book, Bill declined because he felt
group of scientists to develop priorities for mouse that his health precluded his attendance. His
genomicsandgeneticsresources. In responseto the complaintwasthathismemorywasnotasgoodas
community’s recommendations, the NIH created a it used to be. He was 90 at the time. Even so, Bill
Trans-NIH Mouse Genomics and Genetics Re- was invited to give the Keynote Presentation, and
sources Coordinating Group and a strategic imple- thanks to Dabney Johnson (Oak Ridge National
mentationplan,whichincludedinitiativesformouse Laboratory)andLee,filmedavideothatdiscussed
sequencing and database resources, as well as ini- his discovery of ENU, and his reflections on the
tiatives in mouse mutagenesis and phenotyping field to date.
(Batteyetal.,1999).Manyrequestsforapplications Bill’s research program at Oak Ridge National
(RFAs) were issued and funded as a result of the Laboratory was founded just after World War II
NIH response to the recommendations of the com- to study the genetic effects of radiation. The
mittee. The Georgia Genetics Symposium was held developmentof theatomicbomb had ledtomany
prior to the flurry of activities that followed in the concerns about genetic hazards from nuclear fall-
wakeofthisroundofRFAs. out and the continuing use of nuclear energy. Al-
Funding initiatives in mouse genetics in multi- though the hazards of radiation had been
ple countries led to the development of the Inter- established in the fruit fly, the putative parallel in
national Mouse Mutagenesis Consortium mammals, and therefore, the risk to humans, was
(IMMC), established to coordinate mutagenesis not known. In 1947, Bill was asked by Alexander
efforts in the mouse throughout the world. The Hollaender to visit Oak Ridge to develop a re-
goals of the IMMC were published in Science in search program on the genetic effects of radiation
2001, and were to produce at least one heritable in mice. The Biology Division at Oak Ridge Na-
mutation,ineitherEScellsormice,ineverygenein tional Laboratory was a bustling Mecca of genet-
the genome, identify every gene that affects key icistsanddevelopmentalbiologistsworkingwitha
traits of biomedical interest, establish an infra- variety of organisms: among them bacteria, yeast,
structure for preserving and distributing mutant maize, fruit flies, sea urchins, and frogs. The
cells and mice, and enhance the informatics and establishment of Bill’s mouse program was met
database support for these functional studies (Na- with some of the same arguments against large-
deau et al., 2001). In this regard, it is notable that scale mouse mutagenesis today: some argued that
this book contains chapters on nearly each issue the mouse program was unnecessary, since all of
that was subsequentlyproposed bythe IMMC.In the important principles of radiation genetics had
2002, 2 years after the Georgia meeting, the se- been worked out in Drosophila, and others that
quence of the mouse genome was released. Now, genes are genes, and that bacterial genetics would
the draft mouse genome sequence is changing the provide all of the answers (Russell, 1989). Bill ar-
way we approach problems, and is showing that gued that the rate of induction of gene mutations
theseearlymouseinitiativeshavepavedthewayfor in mammals was unknown, and could not be
the mouse to become the leading mammalian ge- extrapolated from the fly. Interestingly, H.J.
netic model organism forthe human. Muller, who won the Nobel Prize for his contri-
butions to the understanding of the genetic effects
of radiation in Drosophila, was one of Bill’s
William L. Russell and the history of ENU greatest supporters in getting his program off the
ground, through funding by the Atomic Energy
Bill Russell of Oak Ridge National Laboratory Commission, later to become the Department of
wasthefatherofmousegermlinemutagenesis,and Energy (DOE).
pioneered the use of the chemical supermutagen Bill’s objective was not simply to measure
N-ethyl-N-nitrosourea (ENU) in mice. Because mutation rate but to investigate the effects of
Athens, Georgia is only a few hours across the physical and biological variables on the quantity
mountainsfromOakRidge,Tennessee,oneofour and the nature of induced mutations. Such data
goals was to have Bill Russell and Lee Russell could only have been obtained by the use of a
attend the meeting. Although Lee obliged, gave a powerful test for the germ line effects of radiation
wonderful talk, and has written an excellent thatBillhadchampioned,calledtheSpecificLocus
5
Test(SLT).Billreminiscedaboutthedevelopment stage, as Bill had done on X-rays using the SLT.
of the SLT for calculating the rate for heritable Finally, in 1978, upon the suggestion of Bill’s
gene mutations induced by radiation or chemicals friend and Drosophila colleague, Ekkehart Vogel,
inthemammaliangermline(forreview,seeDavis Bill carried out a small SLT with ENU, and the
and Justice, 1998). Two of the largest names in rest is history. To his surprise, in the first experi-
genetics at that time, Sewall Wright, Bill’s gradu- ment, this chemical mutagen yielded a mutation
ate mentor, and H.J. Muller, argued that ap- rate 5 times higher than the maximal rate obtain-
proaches other than the SLT would be more able with X-rays in spermatogonia (Russell et al.,
appropriate. Their proposed approaches reflected 1979). By testing various protocols of dose repe-
their experience: Wright suggested measuring tition Russell developed an optimal treatment
longevity and vital statistics in a population of regimen for ENU mutagenesis in male mice that
descendantsofirradiatedmice,whereasMullerfelt yielded mutation rates 12 times higher than
that a specific locus test would be logistically obtainablewithX-rays,andover100timesthatof
impossible in the mouse, and suggested scoring the spontaneous mutation rate, high enough to
visible and lethal mutations over a whole chro- perform forward genetic screens (Hitotsumachi,
mosome. Bill had to remind Muller that mouse Carpenter & Russell, 1985).
geneticslackedsomeoftheelegantgeneticfeatures Although a few pioneering researchers used
of the fly, such as balancer chromosomes that ENU in experiments shortly after its discovery,
would suppress recombination, and that the ap- ENU fell out of favor due to the belief that point
proach would require three generations of breed- mutations induced by ENU would be too difficult
ing instead of one, as in the SLT. Bill won. The to detect molecularly. Bill reflected on the current
specificlocustest‘‘canbecarriedoutsoeasilythat resurgence in forward mutagenesis in the post-
one person, in one hour, without any instrumen- genome era with bittersweet surprise that his dis-
tation, can score [the incidence of gene mutations covery,madeafterhisretirement,wouldhavesuch
in] 2000 loci’’ (Russell, 1989). an impact on the mouse community. To date,
Bill published his initial findings on mutation ENU remains the most powerful germ line muta-
rate in 1951 (Russell, 1951), and publications on gen in the mouse, producing point mutations, ra-
the effects of dose, radiation quality, germ-cell ther than chromosome aberrations. Its use may
stage, and other variables soon followed. A semi- have greater impact in the mouse than point
nalpaperpublishedin1958inScienceshowedthat mutagenesis in the fly. Mouse inbred strains have
protracted radiation to spermatogonia or oocytes exquisitelyuniformgeneticbackgrounds,allowing
(but not to spermatozoa) yielded a considerably for ease of detection of the base pair change.
lower mutation frequency than did acute radia- Along with the genome sequence, as well as im-
tion, and therefore, that mutation rate was influ- proved methodology for single nucleotide poly-
enced by dose rate, in contrast to the findings in morphism detection, finding an ENU-induced
the fruit fly (Russell, Russell & Kelly, 1958). mutation in our database-driven world is
Russell suggested that these cells were capable of straightforward. Through his pioneering work in
repairing premutational damage, thus providing mutagenesis,BillRussellhashelpedustoestablish
oneoftheearliestevidencesfortherepairresearch the mouse as one of the most powerful genetic
that was to follow in many laboratories. organisms today.
WhenBill’sworkbegantoshiftfromradiation
to chemical mutagenesis, one of his goals was to
identify a powerful mouse germ line mutagen; Considerations for mouse mutagenesis
however, the current application of his studies did
not come until after his enforced retirement from Theninechaptersofthisbook,derivedfromtalks
the DOE in 1977. Although more than 20 com- presented at the symposium, cover a variety of
pounds proven to be effective in Drosophila or in topics to consider when creating and analyzing
mammaliansomaticcellshadbeentestedinmouse mouse mutations. Although many mutagenesis
germ cells, and some had been found positive, techniques were presented at the meeting, includ-
none was efficient enough to explore the effect of ing gene trapping and insertional mutagenesis
dose response, dose fractionation, sex and cell using the Sleeping Beauty element, this book