Key Points
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The post-genome challenge is to understand the relationship between our genetic code and physical form and to harness this knowledge to find treatments for disease.
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The mouse is the pre-eminent experimental organism for modelling human disease and determining mammalian gene function.
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Genetics — including the use of genetic screens — offers unsurpassed methods for unravelling complex biological problems.
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Genetic screens fall into two broad categories: region-specific and genome-wide. Within these categories are a host of strategies that can be tailored to suit the aims of individual investigators, regardless of their biological focus.
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Many of the tricks and techniques that have been so useful in lower organisms, such as balancer chromosome and modifier screens, are now being effectively pursued in the mouse.
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Forward genetics in the mouse has been revolutionized since the sequencing of the mouse genome, as the traditional bottleneck — mutation identification — is now relatively simple and straightforward.
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The technical aspects of mouse genetics will continue to become easier; the future is bright for researchers wishing to carry out genetic screens using mice.
Abstract
Humans are mammals, not bacteria or plants, yeast or nematodes, insects or fish. Mice are also mammals, but unlike gorilla and goat, fox and ferret, giraffe and jackal, they are suited perfectly to the laboratory environment and genetic experimentation. In this review, we will summarize the tools, tricks and techniques for executing forward genetic screens in the mouse and argue that this approach is now accessible to most biologists, rather than being the sole domain of large national facilities and specialized genetics laboratories.
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Acknowledgements
Research in the authors' laboratories is supported by a Program Grant from The National Health and Medical Research Council, Canberra, Australia, The Anti-Cancer Council of Victoria, Melbourne, Australia, The J.D. and L. Harris Trust, and The US National Institutes Of Health. B.T.K. is supported by a Queen Elizabeth II Fellowship from the Australian Research Council. We are grateful to N. Nicola, W. Alexander, K. Greig, M. Carpinelli and I. Majewski for critically reading this manuscript.
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Douglas J. Hilton is a founder of the biotechnology company MuriGen, and is a consultant to AMRAD Corporation. Benjamin T. Kile is a consultant to MuriGen.
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DATABASES
Entrez Gene
FURTHER INFORMATION
Children's Hospital Oakland Research Institute BAC/PAC Resources
International Gene Trap Consortium
Jackson Laboratory Mice Database
Jackson Laboratory Mouse Genome Informatics
Jackson Laboratory Mouse Heart, Lung, Blood, and Sleep Disorders Center
Lee Silver's 'Mouse Genetics' online
MIT Center for Genome Research Mouse STS Database
Mouse Gene Prediction Database
Mouse Mutagenesis for Developmental Defects
MRC Mammalian Genetics Unit, Harwell
Mutant Mouse Regional Resource Centers
Oak Ridge National Laboratory — Mammalian Genetics and Genomics
RIKEN (The Institute of Physical and Chemical Research)
Glossary
- MOUSE FANCY
-
A collection of enthusiasts interested in mice with unusual traits.
- FORWARD GENETIC SCREEN
-
A genetic screen in which mutants are isolated on the basis of their phenotype. The mutation responsible is identified by positional cloning or by a candidate-gene approach.
- ALLELIC SERIES
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A series of alleles that are present at the same locus, which produce a gradient of phenotypes.
- SHIRPA
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A hierarchical protocol for the behavioural and pathological phenotyping of potential mouse models of neurological dysfunction.
- EMPReSS
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Individual phenotyping protocols that provide a platform for the systematic and standardized primary screening of mouse mutants.
- EPIGENETIC
-
Any heritable influence (in the progeny of cells or of individuals) on chromosome or gene function that is not accompanied by a change in DNA sequence.
- COMPOUND HETEROZYGOTE
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A diploid genotype in which the two copies of a gene carry different mutations.
- HAPLOINSUFFICIENT
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A gene is haploinsufficient when loss of one functional copy in a diploid organism results in a phenotype.
- HEMIZYGOUS
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A diploid genotype that has only one copy of a particular gene, as in X-chromosome genes in a male, or when the homologous chromosome carries a deletion.
- CRE–LOXP SYSTEM
-
A site-specific recombination system. Two short DNA sequences (loxP sites) are engineered to flank the target DNA. Activation of the Cre recombinase enzyme catalyses recombination between the loxP sites, which leads to the excision of the intervening sequence.
- CONDITIONAL SCREEN
-
A phenotypic screen in which animals, or cells taken from them, are challenged in order to test responses that are not ordinarily detectable in the steady state, for example, by exposure to a chemotherapeutic agent. Both hypo- and hyperresponsive mutants can be recovered.
- HYPOMORPHIC
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A partial loss-of-function allele, sometimes known as a 'weak' or 'leaky' allele.
- THROMBOCYTOPENIA
-
Refers to a reduction in the number of circulating blood platelets.
- ISOGENIC
-
Genetically identical; for example, two mice of the same inbred strain.
- QUANTITATIVE TRAIT
-
A biological trait that shows continuous variation (such as weight) rather than falling into distinct categories (such as diseased or healthy).
- PENETRANCE
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The proportion of affected individuals among the carriers of a particular genotype. If all individuals with a disease genotype show the disease phenotype, then the disease is said to be 'completely penetrant'.
- SIMPLE SEQUENCE LENGTH POLYMORPHISM
-
Simple dinucleotide and trinucleotide repeats that differ in length and, when amplified using PCR, give rise to the generation of fragments of different sizes.
- GRAFT-VERSUS-HOST DISEASE
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A destructive attack on host tissues by immune cells that are derived from a bone marrow transplant.
- SMALL INTERFERING RNAS
-
Small antisense RNAs (20–25 nucleotides long), which are generated from specific dsRNAs, that trigger RNA interference. They serve as guides for the cleavage of homologous mRNA in the RNA-induced silencing complex.
- INVERSE PCR
-
A method for cloning DNA that flanks a known sequence. Genomic DNA is digested and ligated into circles, and is then amplified by PCR. The primers used correspond to the known sequence, but point out from this sequence. In a circle that contains the known sequence, the unknown flanking sequence will be amplified.
- GENE TRAPPING
-
A mutation strategy that uses insertion vectors to trap or isolate transcripts from flanking genes. The inserted sequence functions as a tag from which to clone the mutated gene.
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Kile, B., Hilton, D. The art and design of genetic screens: mouse. Nat Rev Genet 6, 557–567 (2005). https://doi.org/10.1038/nrg1636
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DOI: https://doi.org/10.1038/nrg1636