Genetic Modifier Screens in Mice

Marina R. Carpinelli1, Rachel A. Burt1, Benjamin T. Kile2

1 Murdoch Children's Research Institute, Royal Children's Hospital, Parkville, Australia, 2 Department of Medical Biology, The University of Melbourne, Parkville, Australia
Publication Name:  Current Protocols in Mouse Biology
Unit Number:   
DOI:  10.1002/9780470942390.mo110161
Online Posting Date:  March, 2012
GO TO THE FULL TEXT: PDF or HTML at Wiley Online Library

Abstract

ENU mutagenesis is a forward genetics strategy in which random mutagenesis and phenotypic screening is used to identify genes based on the phenotype induced when they are mutated. A modifier screen is a type of screen in which mice with a pre‐existing phenotype are utilized to identify mutations that can enhance or suppress this phenotype. This approach has the potential to uncover missing pathway members, reveal novel genetic interactions, and pinpoint new drug targets. Considerations when planning a suppressor screen include current knowledge, genomic footprint, penetrance, variance, robustness, latency of the starting phenotype, viability, fertility, genetic background and ENU tolerance of starting strain, screening assay, mouse numbers required, and mutation identification strategy. Practical advice on each of these is provided in this review. Curr. Protoc. Mouse Biol. 2:75‐87 © 2012 by John Wiley & Sons, Inc.

Keywords: ENU mutagenesis; modifier screen; functional genetics

     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Table of Contents

  • Introduction
  • Modifier Screens
  • Summary
  • Literature Cited
  • Figures
  • Tables
     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Materials

GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Figures

Videos

Literature Cited

Literature Cited
   Aigner, B., Rathkolb, B., Mohr, M., Klempt, M., Hrabe de Angelis, M., and Wolf, E. 2007. Generation of ENU‐induced mouse mutants with hypocholesterolemia: Novel tools for dissecting plasma lipoprotein homeostasis. Lipids 42:731‐737.
   Aigner, B., Rathkolb, B., Klaften, M., Sedlmeier, R., Klempt, M., Wagner, S., Michel, D., Mayer, U., Klopstock, T., de Angelis, M.H., and Wolf, E. 2009. Generation of N‐ethyl‐N‐nitrosourea‐induced mouse mutants with deviations in plasma enzyme activities as novel organ‐specific disease models. Exp. Physiol. 94:412‐421.
   Aigner, B., Rathkolb, B., Klempt, M., Wagner, S., Michel, D., Klaften, M., Laufs, J., Schneider, B., Sedlmeier, R., Hrabe de Angelis, M., and Wolf, E. 2011. Generation of N‐ethyl‐N‐nitrosourea‐induced mouse mutants with deviations in hematological parameters. Mamm. Genome 22:495‐505.
   Alexander, W.S., Roberts, A.W., Nicola, N.A., Li, R., and Metcalf, D. 1996. Deficiencies in progenitor cells of multiple hematopoietic lineages and defective megakaryocytopoiesis in mice lacking the thrombopoietic receptor c‐Mpl. Blood 87:2162‐2170.
   Ansorge, W.J. 2009. Next‐generation DNA sequencing techniques. N. Biotechnol 25:195‐203.
   Arnold, C.N., Xia, Y., Lin, P., Ross, C., Schwander, M., Smart, N.G., Muller, U., and Beutler, B. 2011. Rapid identification of a disease allele in mouse through whole genome sequencing and bulk segregation analysis. Genetics 187:633‐641.
   Barbaric, I. and Dear, T.N. 2007. Optimising screening and mating strategies for phenotype‐driven recessive N‐ethyl‐N‐nitrosourea screens in mice. J. Am. Assoc. Lab. Sci. 46:44‐49.
   Barbaric, I., Wells, S., Russ, A., and Dear, T.N. 2007. Spectrum of ENU‐induced mutations in phenotype‐driven and gene‐driven screens in the mouse. Environ. Mol. Mutagen. 48:124‐142.
   Beier, D.R. and Herron, B.J. 2004. Genetic mapping and ENU mutagenesis. Genetica 122:65‐69.
   Beutler, B., Jiang, Z., Georgel, P., Crozat, K., Croker, B., Rutschmann, S., Du, X., and Hoebe, K. 2006. Genetic analysis of host resistance: Toll‐like receptor signaling and immunity at large. Annu. Rev. Immunol. 24:353‐389.
   Bode, V.C. 1984. Ethylnitrosourea mutagenesis and the isolation of mutant alleles for specific genes located in the T region of mouse chromosome 17. Genetics 108:457‐470.
   Bogue, M., Grubb, S., Maddatu, T., and Bult, C. 2007. Mouse phenome database (MPD). Nucleic Acids Res. 35:D643‐D649.
   Brown, S.D., Chambon, P., and de Angelis, M. 2005. EMPReSS: Standardized phenotype screens for functional annotation of the mouse genome. Nat. Genet. 37:1155.
   Brown, S.D., Wurst, W., Kuhn, R., and Hancock, J.M. 2009. The functional annotation of mammalian genomes: The challenge of phenotyping. Annu. Rev. Genet. 43:305‐333.
   Buac, K., Watkins‐Chow, D.E., Loftus, S.K., Larson, D.M., Incao, A., Gibney, G., and Pavan, W.J. 2008. A Sox10 expression screen identifies an amino acid essential for Erbb3 function. PLoS Genet. 4:e100177.
   Carpinelli, M.R., Hilton, D.J., Metcalf, D., Antonchuk, J.L., Hyland, C.D., Mifsud, S.L., Di Rago, L., Hilton, A.A., Willson, T.A., Roberts, A.W., Ramsay, R.G., Nicola, N.A., and Alexander, W.S. 2004. Suppressor screen in Mpl−/− mice: c‐Myb mutation causes supraphysiological production of platelets in the absence of thrombopoietin signaling. Proc. Natl. Acad. Sci. U.S.A. 101:6553‐6558.
   Concepcion, D., Seburn, K.L., Wen, G., Frankel, W.N., and Hamilton, B.A. 2004. Mutation rate and predicted phenotypic target sizes in ethylnitrosourea‐treated mice. Genetics 168:953‐959.
   D'Ascenzo, M., Meacham, C., Kitzman, J., Middle, C., Knight, J., Winer, R., Kukricar, M., Richmond, T., Albert, T.J., Czechanski, A., Donahue, L.R., Affourtit, J., Jeddeloh, J.A., and Reinholdt, L. 2009. Mutation discovery in the mouse using genetically guided array capture and resequencing. Mamm. Genome 20:424‐436.
   Davis, A.P., Woychik, R.P., and Justice, M.J. 1999. Effective chemical mutagenesis in FVB/N mice requires low doses of ethylnitrosourea. Mamm. Genome 10:308‐310.
   de Sauvage, F.J., Hass, P.E., Spencer, S.D., Malloy, B.E., Gurney, A.L., Spencer, S.A., Darbonne, W.C., Henzel, W.J., Wong, S.C., Kuang, W.J. et al. 1994. Stimulation of megakaryocytopoiesis and thrombopoiesis by the c‐Mpl ligand. Nature 369:533‐538.
   Ehling, U.H., Favor, J., Kratochivilova, J., and Neuhauser‐Klaus, A. 1982. Dominant cataract mutations and specific‐locus mutations in mice induced by radiation or ethylnitrosourea. Mutat. Res. 92:181‐192.
   Fairfield, H., Gilbert, G.J., Barter, M., Corrigan, R.R., Curtain, M., Ding, Y., D'Ascenzo, M., Gerhardt, D.J., He, C., Huang, W., Richmond, T., Rowe, L., Probst, F.J., Bergstrom, D.E., Murray, S.A., Bult, C., Richardson, J., Kile, B., Gut, I., Hager, J., Sigurdsson, S., Mauceli, E., Di Palma, F., Lindblad‐Toh, K., Cunningham, M.L., Cox, T.C., Justice, M.J., Spector, M.S., Lowe, S.W., Albert, T., Donahue, L.R., Jeddeloh, J., Shendure, J., and Reinholdt, L.G. 2011. Mutation discovery in mice by whole exome sequencing. Genome Biol. 12:R86.
   Fuchs, H., Gailus‐Durner, V., Adler, T., Aguilar‐Pimentel, J.A., Becker, L., Calzada‐Wack, J., Da Silva‐Buttkus, P., Neff, F., Gotz, A., Hans, W., Holter, S.M., Horsch, M., Kastenmuller, G., Kemter, E., Lengger, C., Maier, H., Matloka, M., Moller, G., Naton, B., Prehn, C., Puk, O., Racz, I., Rathkolb, B., Romisch‐Margl, W., Rozman, J., Wang‐Sattler, R., Schrewe, A., Stoger, C., Tost, M., Adamski, J., Aigner, B., Beckers, J., Behrendt, H., Busch, D.H., Esposito, I., Graw, J., Illig, T., Ivandic, B., Klingenspor, M., Klopstock, T., Kremmer, E., Mempel, M., Neschen, S., Ollert, M., Schulz, H., Suhre, K., Wolf, E., Wurst, W., Zimmer, A., and Hrabe de Angelis, M. 2011. Mouse phenotyping. Methods 53:120‐135.
   Gates, H., Mallon, A.M., and Brown, S.D. 2011. High‐throughput mouse phenotyping. Methods 53:394‐404.
   Ginsburg, D. 2005. Identifying novel genetic determinants of hemostatic balance. J. Thromb. Haemost. 3:1561‐1568.
   Goldsworthy, M., Hugill, A., Freeman, H., Horner, E., Shimomura, K., Bogani, D., Pieles, G., Mijat, V., Arkell, R., Bhattacharya, S., Ashcroft, F.M., and Cox, R.D. 2008. Role of the transcription factor Sox4 in insulin secretion and impaired glucose tolerance. Diabetes 57:2234‐2244.
   Greig, K.T., Antonchuk, J., Metcalf, D., Morgan, P.O., Krebs, D.L., Zhang, J.G., Hacking, D.F., Bode, L., Robb, L., Kranz, C., de Graaf, C., Bahlo, M., Nicola, N.A., Nutt, S.L., Freeze, H.H., Alexander, W.S., Hilton, D.J., and Kile, B.T. 2007. Agm1/Pgm3‐mediated sugar nucleotide synthesis is essential for hematopoiesis and development. Mol. Cell. Biol. 27:5849‐5859.
   Gurney, A.L., Carver‐Moore, K., de Sauvage, F.J., and Moore, M.W. 1994. Thrombocytopenia in c‐mpl‐deficient mice. Science 265:1445‐1447.
   Han, F., Yu, H., Tian, C., Chen, H.E., Benedict‐Alderfer, C., Zheng, Y., Wang, Q., Han, X., and Zheng, Q.Y. 2010. A new mouse mutant of the Cdh23 gene with early‐onset hearing loss facilitates evaluation of otoprotection drugs. Pharmacogenomics J. In press.
   Hardisty‐Hughes, R.E., Parker, A., and Brown, S.D. 2010. A hearing and vestibular phenotyping pipeline to identify mouse mutants with hearing impairment. Nat. Protoc. 5:177‐190.
   Herron, B.J., Lu, W., Rao, C., Liu, S., Peters, H., Bronson, R.T., Justice, M.J., McDonald, J.D., and Beier, D.R. 2002. Efficient generation and mapping of recessive developmental mutations using ENU mutagenesis. Nat. Genet. 30:185‐189.
   Hilton, D.J., Kile, B.T., and Alexander, W.S. 2009. Mutational inhibition of c‐Myb or p300 ameliorates treatment‐induced thrombocytopenia. Blood 113:5599‐5604.
   Hoyne, G.F. and Goodnow, C.C. 2006. The use of genomewide ENU mutagenesis screens to unravel complex mammalian traits: Identifying genes that regulate organ‐specific and systemic autoimmunity. Immunol. Rev. 210:27‐39.
   Johnson, F.M. and Lewis, S.E. 1981. Electrophoretically detected germinal mutations induced in the mouse by ethylnitrosourea. Proc. Natl. Acad. Sci. U.S.A. 78:3138‐3141.
   Johnson, K.R., Zheng, Q.Y., and Erway, L.C. 2000. A major gene affecting age‐related hearing loss is common to at least ten inbred strains of mice. Genomics 70:171‐180.
   Justice, M.J., Noveroske, J.K., Weber, J.S., Zheng, B., and Bradley, A. 1999. Mouse ENU mutagenesis. Hum. Mol. Genet. 8:1955‐1963.
   Justice, M.J., Carpenter, D.A., Favor, J., Neuhauser‐Klaus, A., Hrabe de Angelis, M., Soewarto, D., Moser, A., Cordes, S., Miller, D., Chapman, V., Weber, J.S., Rinchik, E.M., Hunsicker, P.R., Russell, W.L., and Bode, V.C. 2000. Effects of ENU dosage on mouse strains. Mamm. Genome 11:484‐488.
   Kasarskis, A., Manova, K., and Anderson, K.V. 1998. A phenotype‐based screen for embryonic lethal mutations in the mouse. Proc. Natl. Acad. Sci. U.S.A. 95:7485‐7490.
   Kauppi, M., Murphy, J.M., de Graaf, C.A., Hyland, C.D., Greig, K.T., Metcalf, D., Hilton, A.A., Nicola, N.A., Kile, B.T., Hilton, D.J., and Alexander, W.S. 2008. Point mutation in the gene encoding p300 suppresses thrombocytopenia in Mpl‐/‐ mice. Blood 112:3148‐3153.
   Kile, B.T., Hentges, K.E., Clark, A.T., Nakamura, H., Salinger, A.P., Liu, B., Box, N., Stockton, D.W., Johnson, R.L., Behringer, R.R., Bradley, A., and Justice, M.J. 2003. Functional genetic analysis of mouse chromosome 11. Nature 425:81‐86.
   Kimura, S., Roberts, A.W., Metcalf, D., and Alexander, W.S. 1998. Hematopoietic stem cell deficiencies in mice lacking c‐Mpl, the receptor for thrombopoietin. Proc. Natl. Acad. Sci. U.S.A. 95:1195‐1200.
   Levin, J., Cocault, L., Demerens, C., Challier, C., Pauchard, M., Caen, J., and Souyri, M. 2001. Thrombocytopenic c‐mpl(‐/‐) mice can produce a normal level of platelets after administration of 5‐fluorouracil: The effect of age on the response. Blood 98:1019‐1027.
   Lewis, M.A., Quint, E., Glazier, A.M., Fuchs, H., De Angelis, M.H., Langford, C. van Dongen, S., Abreu‐Goodger, C., Piipari, M., Redshaw, N., Dalmay, T., Moreno‐Pelayo, M.A., Enright, A.J., and Steel, K.P. 2009. An ENU‐induced mutation of miR‐96 associated with progressive hearing loss in mice. Nat. Genet. 41:614‐618.
   Loughran, S.J., Kruse, E.A., Hacking, D.F., de Graaf, C.A., Hyland, C.D., Willson, T.A., Henley, K.J., Ellis, S., Voss, A.K., Metcalf, D., Hilton, D.J., Alexander, W.S., and Kile, B.T. 2008. The transcription factor Erg is essential for definitive hematopoiesis and the function of adult hematopoietic stem cells. Nat. Immunol. 9:810‐819.
   Majewski, I.J., Blewitt, M.E., de Graaf, C.A., McManus, E.J., Bahlo, M., Hilton, A.A., Hyland, C.D., Smyth, G.K., Corbin, J.E., Metcalf, D., Alexander, W.S., and Hilton, D.J. 2008. Polycomb repressive complex 2 (PRC2) restricts hematopoietic stem cell activity. PLoS Biol. 6:e93.
   Mamanova, L., Coffey, A.J., Scott, C.E., Kozarewa, I., Turner, E.H., Kumar, A., Howard, E., Shendure, J., and Turner, D.J. 2010. Target‐enrichment strategies for next‐generation sequencing. Nat. Methods 7:111‐118.
   Matera, I., Watkins‐Chow, D.E., Loftus, S.K., Hou, L., Incao, A., Silver, D.L., Rivas, C., Elliott, E.C., Baxter, L.L., and Pavan, W.J. 2008. A sensitized mutagenesis screen identifies Gli3 as a modifier of Sox10 neurocristopathy. Hum. Mol. Genet. 17:2118‐2131.
   Mohan, S., Baylink, D.J., and Srivastava, A.K. 2008. A chemical mutagenesis screen to identify modifier genes that interact with growth hormone and TGF‐beta signaling pathways. Bone 42:388‐395.
   Ng, S.B., Turner, E.H., Robertson, P.D., Flygare, S.D., Bigham, A.W., Lee, C., Shaffer, T., Wong, M., Bhattacharjee, A., Eichler, E.E., Bamshad, M., Nickerson, D.A., and Shendure, J. 2009. Targeted capture and massively parallel sequencing of 12 human exomes. Nature 461:272‐276.
   Nijman, I.J., Mokry, M., van Boxtel, R., Toonen, P., de Bruijn, E., and Cuppen, E. 2010. Mutation discovery by targeted genomic enrichment of multiplexed barcoded samples. Nat. Methods 7:913‐915.
   Noben‐Trauth, K., Zheng, Q.Y., and Johnson, K.R. 2003. Association of cadherin 23 with polygenic inheritance and genetic modification of sensorineural hearing loss. Nat. Genet. 35:21‐23.
   Pyrgaki, C., Liu, A., and Niswander, L. 2011. Grainyhead‐like 2 regulates neural tube closure and adhesion molecule expression during neural fold fusion. Dev. Biol. 353:38‐49.
   Quwailid, M.M., Hugill, A., Dear, N., Vizor, L., Wells, S., Horner, E., Fuller, S., Weedon, J., McMath, H., Woodman, P., Edwards, D., Campbell, D., Rodger, S., Carey, J., Roberts, A., Glenister, P., Lalanne, Z., Parkinson, N., Coghill, E.L., McKeone, R., Cox, S., Willan, J., Greenfield, A., Keays, D., Brady, S., Spurr, N., Gray, I., Hunter, J., Brown, S.D., and Cox, R.D. 2004. A gene‐driven ENU‐based approach to generating an allelic series in any gene. Mamm. Genome 15:585‐591.
   Rogers, D.C., Fisher, E.M., Brown, S.D., Peters, J., Hunter, A.J., and Martin, J.E. 1997. Behavioral and functional analysis of mouse phenotype: SHIRPA, a proposed protocol for comprehensive phenotype assessment. Mamm. Genome 8:711‐713.
   Rubio‐Aliaga, I., Soewarto, D., Wagner, S., Klaften, M., Fuchs, H., Kalaydjiev, S., Busch, D.H., Klempt, M., Rathkolb, B., Wolf, E., Abe, K., Zeiser, S., Przemeck, G.K., Beckers, J., and de Angelis, M.H. 2007. A genetic screen for modifiers of the delta1‐dependent notch signaling function in the mouse. Genetics 175:1451‐1463.
   Russell, W.L., Kelly, E.M., Hunsicker, P.R., Bangham, J.W., Maddux, S.C., and Phipps, E.L. 1979. Specific‐locus test shows ethylnitrosourea to be the most potent mutagen in the mouse. Proc. Natl. Acad. Sci. U.S.A. 76:5818‐5819.
   Sandberg, M.L., Sutton, S.E., Pletcher, M.T., Wiltshire, T., Tarantino, L.M., Hogenesch, J.B., and Cooke, M.P. 2005. c‐Myb and p300 regulate hematopoietic stem cell proliferation and differentiation. Dev. Cell 8:153‐166.
   Sheridan, R., Lampe, K., Shanmukhappa, S.K., Putnam, P., Keddache, M., Divanovic, S., Bezerra, J., and Hoebe, K. 2011. Lampe1: An ENU‐germline mutation causing spontaneous hepatosteatosis identified through targeted exon‐enrichment and next‐generation sequencing. PLoS One 6:e21979.
   Silver, J.D., Hilton, D.J., Bahlo, M., and Kile, B.T. 2007. Probabilistic analysis of recessive mutagenesis screen strategies. Mamm. Genome 18:5‐22.
   Silver, L.M. 1995. Mouse Genetics. Oxford University Press, New York.
   Smith, A.P., Polley, S., Wells, S., Stewart, M., Vizor, L., Humphreys, J., Warren, M.V., Dear, N., and Cheeseman, M.T. 2009. Analysis of breeding and pathology helps refine management practices of a large‐scale N′‐ethyl‐N′‐nitrosourea mouse mutagenesis programme. Lab. Anim. 43:1‐10.
   Speca, D.J., Rabbee, N., Chihara, D., Speed, T.P., and Peterson, A.S. 2006. A genetic screen for behavioral mutations that perturb dopaminergic homeostasis in mice. Genes Brain Behav. 5:19‐28.
   Stottmann, R.W., Moran, J.L., Turbe‐Doan, A., Driver, E., Kelley, M., and Beier, D.R. 2011. Focusing forward genetics: A tripartite ENU screen for neurodevelopmental mutations in the mouse. Genetics 188:615‐624.
   Su, L.K., Kinzler, K.W., Vogelstein, B., Preisinger, A.C., Moser, A.R., Luongo, C., Gould, K.A., and Dove, W.F. 1992. Multiple intestinal neoplasia caused by a mutation in the murine homolog of the APC gene. Science 256:668‐670.
   Tchekneva, E.E., Rinchik, E.M., Polosukhina, D., Davis, L.S., Kadkina, V., Mohamed, Y., Dunn, S.R., Sharma, K., Qi, Z., Fogo, A.B., and Breyer, M.D. 2007. A sensitized screen of N‐ethyl‐N‐nitrosourea‐mutagenized mice identifies dominant mutants predisposed to diabetic nephropathy. J. Am. Soc. Nephrol. 18:103‐112.
   Vitaterna, M.H., King, D.P., Chang, A.M., Kornhauser, J.M., Lowrey, P.L., McDonald, J.D., Dove, W.F., Pinto, L.H., Turek, F.W., and Takahashi, J.S. 1994. Mutagenesis and mapping of a mouse gene, clock, essential for circadian behavior. Science 264:719‐725.
   Wansleeben, C., van Gurp, L., Feitsma, H., Kroon, C., Rieter, E., Verberne, M., Guryev, V., Cuppen, E., and Meijlink, F. 2011. An ENU‐mutagenesis screen in the mouse: Identification of novel developmental gene functions. PLoS One 6:e19357.
   Waterston, R.H., Lindblad‐Toh, K., Birney, E., Rogers, J., Abril, J.F., Agarwal, P., Agarwala, R., Ainscough, R., Alexandersson, M., An, P., Antonarakis, S.E., Attwood, J., Baertsch, R., Bailey, J., Barlow, K., Beck, S., Berry, E., Birren, B., Bloom, T., Bork, P., Botcherby, M., Bray, N., Brent, M.R., Brown, D.G., Brown, S.D., Bult, C., Burton, J., Butler, J., Campbell, R.D., Carninci, P., Cawley, S., Chiaromonte, F., Chinwalla, A.T., Church, D.M., Clamp, M., Clee, C., Collins, F.S., Cook, L.L., Copley, R.R., Coulson, A., Couronne, O., Cuff, J., Curwen, V., Cutts, T., Daly, M., David, R., Davies, J., Delehaunty, K.D., Deri, J., Dermitzakis, E.T., Dewey, C., Dickens, N.J., Diekhans, M., Dodge, S., Dubchak, I., Dunn, D.M., Eddy, S.R., Elnitski, L., Emes, R.D., Eswara, P., Eyras, E., Felsenfeld, A., Fewell, G.A., Flicek, P., Foley, K., Frankel, W.N., Fulton, L.A., Fulton, R.S., Furey, T.S., Gage, D., Gibbs, R.A., Glusman, G., Gnerre, S., Goldman, N., Goodstadt, L., Grafham, D., Graves, T.A., Green, E.D., Gregory, S., Guigo, R., Guyer, M., Hardison, R.C., Haussler, D., Hayashizaki, Y., Hillier, L.W., Hinrichs, A., Hlavina, W., Holzer, T., Hsu, F., Hua, A., Hubbard, T., Hunt, A., Jackson, I., Jaffe, D.B., Johnson, L.S., Jones, M., Jones, T.A., Joy, A., Kamal, M., Karlsson, E.K., Karolchik, D., Kasprzyk, A., Kawai, J., Keibler, E., Kells, C., Kent, W.J., Kirby, A., Kolbe, D.L., Korf, I., Kucherlapati, R.S., Kulbokas, E.J., Kulp, D., Landers, T., Leger, J.P., Leonard, S., Letunic, I., Levine, R., Li, J., Li, M., Lloyd, C., Lucas, S., Ma, B., Maglott, D.R., Mardis, E.R., Matthews, L., Mauceli, E., Mayer, J.H., McCarthy, M., McCombie, W.R., McLaren, S., McLay, K., McPherson, J.D., Meldrim, J., Meredith, B., Mesirov, J.P., Miller, W., Miner, T.L., Mongin, E., Montgomery, K.T., Morgan, M., Mott, R., Mullikin, J.C., Muzny, D.M., Nash, W.E., Nelson, J.O., Nhan, M.N., Nicol, R., Ning, Z., Nusbaum, C., O'Connor, M.J., Okazaki, Y., Oliver, K., Overton‐Larty, E., Pachter, L., Parra, G., Pepin, K.H., Peterson, J., Pevzner, P., Plumb, R., Pohl, C.S., Poliakov, A., Ponce, T.C., Ponting, C.P., Potter, S., Quail, M., Reymond, A., Roe, B.A., Roskin, K.M., Rubin, E.M., Rust, A.G., Santos, R., Sapojnikov, V., Schultz, B., Schultz, J., Schwartz, M.S., Schwartz, S., Scott, C., Seaman, S., Searle, S., Sharpe, T., Sheridan, A., Shownkeen, R., Sims, S., Singer, J.B., Slater, G., Smit, A., Smith, D.R., Spencer, B., Stabenau, A., Stange‐Thomann, N., Sugnet, C., Suyama, M., Tesler, G., Thompson, J., Torrents, D., Trevaskis, E., Tromp, J., Ucla, C., Ureta‐Vidal, A., Vinson, J.P., Von Niederhausern, A.C., Wade, C.M., Wall, M., Weber, R.J., Weiss, R.B., Wendl, M.C., West, A.P., Wetterstrand, K., Wheeler, R., Whelan, S., Wierzbowski, J., Willey, D., Williams, S., Wilson, R.K., Winter, E., Worley, K.C., Wyman, D., Yang, S., Yang, S.P., Zdobnov, E.M., Zody, M.C., and Lander, E.S. 2002. Initial sequencing and comparative analysis of the mouse genome. Nature 420:520‐562.
   Weber, J.S., Salinger, A., and Justice, M.J. 2000. Optimal N‐ethyl‐N‐nitrosourea (ENU) doses for inbred mouse strains. Genesis 26:230‐233.
   Westrick, R.J. and Ginsburg, D. 2009. Modifier genes for disorders of thrombosis and hemostasis. J. Thromb. Haemost. 7:132‐135.
   Wilkie, A.O. 1994. The molecular basis of genetic dominance. J. Med. Genet. 31:89‐98.
   Xia, Y., Won, S., Du, X., Lin, P., Ross, C., La Vine, D., Wiltshire, S., Leiva, G., Vidal, S.M., Whittle, B., Goodnow, C.C., Koziol, J., EM, Y.M., and Beutler, B. 2010. Bulk segregation mapping of mutations in closely related strains of mice. Genetics 186:1139‐1146.
   Yabas, M., Teh, C.E., Frankenreiter, S., Lal, D., Roots, C.M., Whittle, B., Andrews, D.T., Zhang, Y., Teoh, N.C., Sprent, J., Tze, L.E., Kucharska, E.M., Kofler, J., Farell, G.C., Broer, S., Goodnow, C.C., and Enders, A. 2011. ATP11C is critical for the internalization of phosphatidylserine and differentiation of B lymphocytes. Nat. Immunol. 12:441‐449.
   Zhang, Z., Alpert, D., Francis, R., Chatterjee, B., Yu, Q., Tansey, T., Sabol, S.L., Cui, C., Bai, Y., Koriabine, M., Yoshinaga, Y., Cheng, J.F., Chen, F., Martin, J., Schackwitz, W., Gunn, T.M., Kramer, K.L., De Jong, P.J., Pennacchio, L.A., and Lo, C.W. 2009. Massively parallel sequencing identifies the gene Megf8 with ENU‐induced mutation causing heterotaxy. Proc. Natl. Acad. Sci. U.S.A. 106:3219‐3224.
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library