Mouse Strains and Resources Introduction Mouse Strains and Resources The tcigon fees at ey Welcome to Inbred Mouse Strains and Resources. Inbred mouse strains are commonly used in genetic and genomic studies here at The Jackson Laboratory. As you may know, there is more than one inbred strain of mouse. In fact, there are hundreds of in-bred strains of mice, some of which may be really beneficial for your studies. And some of which you'll want to avoid. Learning Outcomes eT) eee eee eee Te Creer ard erates rbd No audio The Jackson LaboratoryInbred Strains Mouse as a Model for Understanding Human Biology and Disease eee cece Click RIGHT ARROW to continue Why are mice such a useful model for genetic and genomic research? Genetically Similar Mouse as a Model for Understanding Human Biology and Disease Genetically Similar: Certainly mice and humans are genetically very similar; we share between 95 and 98 percent of our genomes. The Jackson LaboratoryPhysiologically Anatomically Similar Ee can and isease Physiologically and anatomically simitar i fe eet hall Physiologically and Anatomically Similar: Besides genetic similarity, we are also physiologically and anatomically similar. In this illustration we see that a similarity in the respiratory system enables us to model human lung cancer in mice. Also, humans and mice share many of the same neuromuscular diseases. In fact, the same genetic causes of, for example, muscular dystrophy and motor neuron diseases, apply to both humans and mice because the basic anatomical “wiring” and functions are so very similar. Experimental Modification of Mouse Genes: Mouse as a Model for Understanding Human Biology and Disease regi yi he cca ~ Come Experimental Modification of Mouse Genes: We can do some amazing things working with mice. We can genetically modify our mice in numerous ways, and now with CRISPR technology we can make very precise genetic changes. We can make transgenic mice (adding genes] and we can make knock outs (removing genes). And we can make conditional knock outs. We can put human genes in, We can even rearrange chromosomes. All these genetic modifications can make your mouse a better model for whatever disease or whatever biological process you want to study. SX The Jackson LaboratoryGenetic Modifiers Mouse as a Model for Understanding Human Biology and Disease Ability to study genetic modifiers Click RIGHT ARROW to continue Genetic Modifiers: We can also study genetic modifiers in our mice. Although genetic modifiers show up all the time in human pedigrees, they're really hard to study because there never are enough segregating phenotypes in a family to really do the genetics appropriately, making appropriate genetic analyses impossible. Sometimes it’s hard enough to do the Mendelian genetics, and then when you start adding multiple genes on top of that, it gets very complex, very quickly. Here at The Jackson Laboratory we breed many mice, and you can actually study not only the initial disease causing gene, but you can also mix the genomes from inbred strains with very different origins to allow mapping and identification of genetic modifiers. Even in these ideal circumstances, the segregation of more than two or three genes makes for “complex trait” genetics! Laboratory Mouse Strains ETO Atos eT Classical inbred Originally laboratory mouse strains were generated from fancy mice which were kept The Jackson Laboratoryas pets. They captured alleles and phenotypes from ancestral wild mouse subspecies and were subsequently selected to represent interesting mutant phenotypes. The wild type alleles (from the ancestral wild mice] often exert dominance over most spontaneous mutations; therefore, “fancy” mutations must be maintained carefully to preserve the desired phenotypes. Inbred Strains Inbred Strains Outbred, or randomly bred mice, possess unpredictable genetic variation and many heterozygous loci, whereas inbred mouse strains are genetically identical and homozygous at every locus. The scientific world as a whole, and specifically the field of biomedical research, has benefited incredibly from inbred strains of mice. There aren't very many animal model organisms that you can study where you're actually working with a population of genetically identical individuals. Many investigators can simultaneously work on these genetically identical mice and compare results, making it an amazing resource. Types of Inbred Strains Types of Inbred Strains @ sre @ Wissserives Mae muscu domestne as mused muscle PHI ve muscu rsa ‘Mas mucus castneus EASE There are two types of inbred strain, standard and wild-derived. The Jackson LaboratoryStandard laboratory strains were derived from pet mice maintained by mouse fanciers in Europe and Asia and that contained genetic contributions from Mus musculus domesticus, musculus, molossinus and castaneus. Most of the inbred strains started out as pet store mice that were then inbred. When you domesticate any species, you eliminate wild characteristics as well, by selecting for those animals that are easy to handle and easy to breed. The disadvantage is that you've now also selected against lots of geneticvariation. Because of this, scientists have found it useful to outcross mice from a standard inbred strain with a wild-derived strain in genetic mapping studies wherein the wild-derived genome introduces thousands of variant genetic markers. Wild-derived strains are derived from a single subspecies or population. They provide a reagent that is genetically distinct from standard inbred strains. Examples include ‘Mus musculus musculus, Mus musculus castaneus, and Mus spretus. Mus spretus is. actually a different species all together, where F1 hybrid matings with common lab strains produce males that are sterile and females that are fertile. Genetically, they're very interesting, but frequently difficult to handle because they still exhibit wild behavior. They are often referred to as “popcorn mice” because of their propensity to fly out of cages as soon as the cage lid is opened. They also are savage little biters! Standard Inbred Strains EERIE CS7BL/6s MT As Sead Inbred strains have unique characteristics due to the genetic variants and mutations that were fixed and selected into the background during inbreeding. In many cases, these traits were selected for during the inbreeding process. These are not flaws, rather they are features. These are examples of inbred mouse strains we commonly use for genetic and genomic research, starting with the C57BL/6J; the most widely used inbred strain. As you will see, each strain has its own characteristics that determine whether it is an ideal or less The Jackson Laboratorythan ideal mouse model for your research. C57BL/6J Standard Inbred Strains permssie backround for maximal expression of most main. 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The ar ero ec Vid tan, tran ‘racer of akRiac couse alterations nha devsopmer ate ip edent croscopy Aeracrtal i deletion all 9 KR ‘tice iscausedbyamuston insert Oaoftrenterace Satan (Gade toetraneated OAT! protein AMR mice ar ralabvely rei 6 sorties oration onasem-sntetichigh at tanger Iiporesponsiete diets corning gh levels offal chalesterk lek etter itoration The Jackson LaboratoryStandard Inbred Strains {cle ttslesons have ben Trequener Some CBA! mice sponaneast develo exci panei Ineleney jeome lek ett oration DBA/2J TEER UCU CLI attic a widely ucedinired train Some characteris inl le ‘cypityt developing ateroslrtc orc eslon, ah trequencyhearng los sucepbiiy te selopene tours, trvelopnet of progressive ee orm tat coal aan ereiary glaucoma sndestrame ntleance oss ane rerpine lek ett oration C3H/HeJ Standard Inbred Strains encores tod ‘maion ocean {Cian popeteccrie response acs (ation ntl ecegtr£ gee, r=" maxing 3H Med mice mare reset to ‘aston Cie Tet maze are gny sarepletoieson by ram nae scar seh ae Simones tric. lek ett intonation ANS The Jackson LaboratoryCRC LIS Fr ge AND {Ris highly susceptibe to cortsone-indoced congerstal ct pala has CoA | Satopertnct tran thppeccene tec penitent ae aera tl aon ryote cls lari oxcrine nds havebeenoberved in The lek ett oration Inbred Breeding Scheme TL) tranedual norcayaoety t ae orate, Nrneammnnste ‘goneratlons of Inbreeding ver 195 Inbred mice result from at least 20 sequential generations of brother-sister mating. After 20 generations of inbreeding, the mouse has reached the fully inbred status. In an inbred population, the progeny are now genetically identical. Since the inbred mouse is reproducible, we can study exactly the same organism at The Jackson Laboratory in Bar Harbor as in California, Germany, or Australia. Also, the inbred mouse you are studying today can be compared to that contributing measurement made five yearsago. Inbreeding allows you to do repeated measurements, particularly useful for phenotypes that tend to be subtle. Most of our inbred mouse strains at The Jackson Laboratory have been inbred for hundreds of generations - many hundreds, in some cases. At 20 generations, the inbred mouse is 98.6% homozygous. However, most of our inbred strains at the lab are hundreds of generations inbred and are closer to 99.999% homozygous. Genetic Drift The Jackson LaboratoryvB CC sent 7 eee 3 ee in By We ‘ereratons nae Ifyou start managing your own colony of inbred mice, it very rapidly can become your own substrain, indicating that it may no longer be genetically and phenotypically equivalent to the source colony. This is due to genetic drift that results from accumulated random mutations as inbreeding continues away from the source colony. So, in colony maintenance you will always want to refresh your breeding colony every five generations with mice from the standardized source colony such as The Jackson Laboratory where we closely monitor the genetics of our inbred strains. Phenotype Variation among Laboratory Mouse Strains eM 5 ATC) i eae OAT. Ee I CECE Ear : i Laboratory mouse strains differ genetically and phenotypically. The Jackson Laboratory maintains both the Mouse Phenome Database and the Mouse Genome Informatics (MGIl database. It serves as a primary resource for a spectrum of genetic, genomic and biological data supporting the use of the mouse as a model for understanding human biology and disease. As inbred mice differ widely in characteristics among each other, you can search these strain databases for any kind of phenotype you're interested in, including measurements of anatomy, behavior, blood, cancer, diet effects, drug effects, genotype, heart/lung, metabolic, neurosensory, reproduction and skeletal parameters. The Jackson LaboratoryThis diagram illustrates the large amount of variation that exists for the trait that is being measured among the strains listed. The Mouse Phenome Database is an incredibly useful tool to compare data. It has information from over 250 strains, and over 4000 phenotypes -- that have been assessed. It also has sequencing information containing millions of SNPs throughout the entire genome that have been mapped to these different strains. Self Check! on Consider the following statement: SEI) Fiat matings are required for atleast 20 generations to creat the gonetiealy homogeneous inbred strains valuable for biomedical research @ me False rE No audio Mating Schemes Mating Schemes. toed uterase Inow strains 1 hybrids mapping. ongeis) azn) murs nana) AlnsAls The Jackson LaboratoryThere are several types of mating schemes that can be performed to maintain your colonies and to answer your experimental question. Incross Ifyou are maintaining an inbred line, you are performing an incross mating scheme. You are mating two animals that are homozygous for the same genotype at any designated locus. Since both parents are homozygous, their offspring are also going to be homozygous for the same alleles. Outcross An outcross is simply a crossbetween two different strains that are homozygous for a different genetic sequence at any locus where these strains differ. Normally these individuals are unrelated and their progeny will be F1 hybrids, the first filial generation, heterozygous at every locus across the whole genome. One important feature about the F1 hybrids between inbred strains is that they are all identical to one another. But once you breed the F1 hybrids, now they will segregate the different alleles from the two inbred strains to their offspring, which therefore will not be identical to each other. Backcross A backcross utilizes the F1 hybrids that we've just generated here, and crosses them back to one of the parental inbred strains. The result is segregation across the genome, but at any one locus, 50% of the mice will be homozygous for the backcross parental genotype, and 50% of the mice will be heterozygous. Intercross When you take two F1 hybrid mice that have the same heterozygous genotype across the genome, and cross them together, you are performing an intercross. If the two parental strains carry a different variant of the same gene [allele], then the F2 population shows the Mendelian pattern of 25% homozygous for either strain allele and 50% heterozygous. Genetic Resources COT Ce ce cack Coisogenies Recombinant Inbreds Congenics Coreen Together with the hundreds of inbred mouse strains, classic breeding strategies can be The Jackson Laboratoryand are utilized to make new genetic resources. Coisogenics COT Cea ce cucky CConsomics SE (crc | aoe” =z ae yn atonly a single locus. Coisogenic: A coisogenic strain is a variant strain of mice that differs from an established inbred strain by a mutation at only a single locus. If a new mutation has arisen on that inbred strain, the whole genome is still like the parental type, except for the one locus where the mutation occurred. Coisogenic strains are used in phenotype driven research to identify genes or the function of a gene. Example Coisogenics The Jackson LaboratoryCOT Ce ce cack Coisogenies osreL/e.a Recombinant Inbreds Consomics ) (e57BL/6-tub/tud Congenics C5TBLIS-o0/00 Example Coisogeni These mice are all coisogenic; they are from C57BL/6J parent of origin with new mutations: a yellow agouti, a Tubby mutation and an obese mutation. They are exactly like the original C57BL/6J except for the single place where the mutation occurs in the genome . Recombinant Inbred COT Cea ce cucky Coisogenics: Recombinant Inbred ar ar Recombinant wezarome UIT JT oe eS Congenics. Fis gees que , qu “I Sibling matings for > 20, Recombinant Inbred: Recombinant inbred strains are a special type of inbred strain formed from an initial outcross between two well-characterized inbred strains followed by at least 20, generations of inbreeding. There will be a random variety of allele combinations across the chromosomes and across individuals in the F2 generation. Recombinant inbreds are used as mapping reagents; a strain distribution pattern of any particular phenotype can be compared with genotypes that have previously been mapped in the Recombinant Inbred set. They are particularly useful when mapping quantitative traits with large inter-individual variability. The Jackson LaboratoryConsomics COT Ce ce cack Coisogenics Consomics: ar 4 ae | sesame NN = Inbred strains — Congenics backerosses for entire chromosome followed by intercross. qi Consomic: Aconsomic strain is a kind of strain that contains a single entire chromosome from another strain using a backcross breeding scheme. First, two different strains are mated, and then one chromosome is selected for by using genetic markers in the offspring at each backcross generation. After at least 10 successive backcrosses, the selected chromosome is made homozygous by a final intercross. Aconsomic is used if you would like to rapidly associate a phenotype with a particular chromosome for which the donor and the host differ. This is useful when analyzing complex traits or identifying genetic modifiers. Because the consomic stock introduces thousands of genetic variants into the recipient, it is usually only the first step in identifying the role of a given chromosome in control of the phenotype of interest. Congenics COT Ce ce cack Congenies a Wt seasesst QT = YT seem NM Sactmnce’— |N Congenic: Congenics use similar backcrossing schemes as consomics and result in the replacement of a small region of a single chromosome with DNA from another strain. A congenic strain is the same as the parental inbred strain except for the one piece of The Jackson Laboratorythe genome you transferred from another strain. After at least ten backcross generations and selecting for the region of interest using genetic markers, additional markers spread across the genome can be used to confirm that the resulting congenic strain is 99.9 percent derived from the backcross parent. Self Check! REM cot cncossoecn No Audio End of Presentation No audio The Jackson Laboratory