Page 54 - Mouse Molecular Genetics

Full Abstracts
Program number is above title. Author in bold is the presenter.
54
long term humoral immunity. I will also discuss diversity of helper T cells specialized in humoral immunity, and show that a
subpopulation of these cells are localized in germinal centers by sharing trafficking molecules with germinal center B cells.
Genetics and Genomics
57
KOMP2...the next phase of the Knockout Mouse Project. Kent Lloyd
.
Mouse Biology Program, Univ California Davis,
Davis, CA.
As part of KOMP2, an NIH Common Fund project to produce and phenotype The Knockout Mouse Production and
Phenotyping (KOMP2) project is an NIH Common Fund program intended to conduct broad based, comprehensive, high
throughput phenotyping on homozygous mutant mouse lines produced from gene targeted embryonic stem (ES) cells generated
during the last 5 years of KOMP1. Three KOMP2 production and phenotyping centers were awarded to 3 groups: the DTCC
Consortium led by Kent Lloyd at UC Davis and including the Toronto Center for Phenogenomics, Childrens Hospital Oakland
Research Institute, and Charles River Laboratory; the BaSH Consortium led by Monica Justice at Baylor College of Medicine
and including The Sanger Institute and Harwell; and The Jackson Laboratory led by Robert Braun and Leah Donohue. In
addition, a Knockout Mouse Phenotyping Project Database was awarded to Dr. Paul Flicek at the European Bioinformatics
Institute and will create a data coordination center to collect, archive, and provide online access via a web portal phenotyping
data and information generated from the 3 KOMP2 award recipients, as well as complex queries and statistical analyses of the
data. Over the next 5 years, for 2500, mostly unannotated genes, live mice, cryopreserved germplasm, and an abundance of data
will be generated from a phenotyping pipeline covering all major body systems and most areas of human disease. KOMP2 will
provide a unique and valuable scientific resource for the biomedical research community, and will inform our understanding of
the pathophysiology of human and animal diseases. If successful, another 5000 genes will be analyzed over an additional 5 years.
The KOMP2 efforts are being coordinated with complementary international efforts. The KOMP2 project awarded to the DTCC
is supported by NIH grants U42 OD011175 and U54HG006364.
58
High-throughput mutant mouse phenotyping is a powerful tool to generate novel hypotheses. Chris Lelliott
,
Jeanne
Estabel, Anna Karin Gerdin, Antonella Galli, Anneliese Speak, Richard Houghton, Joanna Bottomley, Edward Ryder, Ramiro
Ramirez-Solis, David Adams, Jacqueline White. The Sanger Mouse Genetics Project, Wellcome Trust Sanger Institute, Hinxton,
Cambridgeshire, United Kingdom.
As part of the Sanger Mouse Genetics Project we have generated over 600 knockout mouse lines. Viability and fertility of each
line are assessed during colony expansion; 28% of lines have been found to be embryonic lethal at postnatal day 14, with a
further 12% classed as sub-viable, and 5.7% of lines homozygous for the targeted allele present with fertility issues. Following
expansion, dedicated groups of mice are phenotyped through an extensive and standardised battery of tests assessing
development, physiology and metabolism, and yielding data on 280 parameters for every line. For homozygous lethal lines,
heterozygous animals are phenotyped. Whilst lines homozygous for the targeted alleles present with a higher hit rate (64% of
lines assessed in the homozygous state presented with 1 hit and averaged 3.9 hits per line), lines assessed in the heterozygous
state were still remarkably informative (42% of lines assessed in the heterozygous state were classed as pheno-deviant with an
average of 1 hit per line) and represent a rich source of information, which is openly available on the Sanger Mouse Portal
A significant proportion of genes have been selected based on the absence of available
mutant mouse phenotypic information. We will present some of the hypothesis-generating mouse lines that have resulted,
including mice with metabolic abnormalities (
Kptn
,
Dusp3
),
skeletal and behavioural changes (
Zc3hc1
),
haematological
alterations (
Crlf3
)
and developmental defects (
Psat1
).
Our data shows that high-throughput phenotyping is a powerful tool to
generate novel hypotheses and that most knockouts have robust phenotypes, few of which could be predicted a
priori
.
59
Recent specialization of the human and mouse X chromosomes for the male germline. Jacob L. Mueller
1
,
Helen Skaletsky
1
,
Laura G. Brown
1
,
Sara Zaghlul
1
,
Susan Rock
2
,
Tina Graves
2
,
Katherine Auger
3
,
Wesley C. Warren
2
,
Richard K. Wilson
2
,
David
C. Page
1
. 1) 1
Howard Hughes Medical Institute, Whitehead Institute, MIT, Cambridge, MA; 2) The Genome Institute,
Washington University School of Medicine, St. Louis, MO; 3) The Wellcome Trust Sanger Institute, Cambridge, United
Kingdom.
In 1967, Susumu Ohno posited that the gene content of X chromosomes would be conserved across placental mammals, a
statement often referred to as Ohnos law. This widely accepted law has not yet been tested systematically and rigorously. We
decided to test Ohnos law by comparing the human X chromosome to the single-haplotype, clone-based, mouse X-chromosome
sequence. This required that we refine portions of the current human X-chromosome reference sequence with single-haplotype
sequence to match the single-haplotype mouse sequence. Single-haplotype, clone-based sequence was selectively generated on
large, nearly identical, segmental duplications, termed amplicons. We resolved gaps in the current reference sequence,