Abstract
In 2007 Capecchi, Evans, and Smithies received the Nobel Prize in recognition for discovering the principles for introducing specific gene modifications in mice via embryonic stem cells, a technology, which has revolutionized the field of biomedical science allowing for the generation of genetically engineered animals. Here we describe detailed protocols based on and developed from these ground-breaking discoveries, allowing for the modification of genes not only to create mutations to study gene function but additionally to modify genes with fluorescent markers, thus permitting the isolation of specific rare wild-type and mutant cell types for further detailed analysis at the biochemical, pathological, and genomic levels.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Ghigna C, Goodell J (1999) Mice Are Nice. Random House Children’s Books. ISBN: 9780679989295
Skarnes WC, Rosen B, West AP et al (2011) A conditional knockout resource for the genome-wide study of mouse gene function. Nature 474:337–342
Bower DV, Sato Y, Lansford R (2011) Dynamic lineage analysis of embryonic morphogenesis using transgenic quail and 4D multispectral imaging. Genesis 49:619–643
Day RN, Davidson MW (2009) The fluorescent protein palette: tools for cellular imaging. Chem Soc Rev 38:2887–2921
Hadjantonakis AK, Nagy A (2001) The color of mice: in the light of GFP-variant reporters. Histochem Cell Biol 115:49–58
Shaner NC, Steinbach PA, Tsien RY (2005) A guide to choosing fluorescent proteins. Nat Methods 2:905–909
Chan HY, V S, Xing X et al. (2011) Comparison of IRES and F2A-based locus-specific multicistronic expression in stable mouse lines. PLoS One 6:e28885
Feil R (2007) Conditional somatic mutagenesis in the mouse using site-specific recombinases. Handb Exp Pharmacol (178) 3–28
Kraus P, Sivakamasundari V, Lim SL et al (2013) Making sense of Dlx1 antisense RNA. Dev Biol 376:224–235
Lee WJ, Kraus P, Lufkin T (2012) Endogenous tagging of the murine transcription factor Sox5 with hemagglutinin for functional studies. Transgenic Res 21:293–301
Sivakamasundari V, Chan HY, Yap SP et al (2012) New Bapx1(Cre-EGFP) mouse lines for lineage tracing and conditional knockout studies. Genesis 50:375–383
Sivakamasundari V, Kraus P, Jie S et al (2013) Pax1(EGFP): New wild-type and mutant EGFP mouse lines for molecular and fate mapping studies. Genesis 51(6):420–429
Yap SP, Xing X, Kraus P et al (2011) Generation of mice with a novel conditional null allele of the Sox9 gene. Biotechnol Lett 33:1551–1558
Kraus P, Xing X, Lim SL et al (2012) Mouse strain specific gene expression differences for illumina microarray expression profiling in embryos. BMC Res Notes 5:232
Nagy A, Getsenstein M, Vintersten K et al (2002) Manipulating the mouse embryo: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor
Soriano P (1999) Generalized lacZ expression with the ROSA26 Cre reporter strain. Nat Genet 21:70–71
Metzger D, Chambon P (2001) Site- and time-specific gene targeting in the mouse. Methods 24:71–80
Metzger D, Clifford J, Chiba H et al (1995) Conditional site-specific recombination in mammalian cells using a ligand-dependent chimeric Cre recombinase. Proc Natl Acad Sci U S A 92:6991–6995
Zhang J, Zhao J, Jiang WJ et al (2012) Conditional gene manipulation: Cre-ating a new biological era. J Zhejiang Univ Sci B 13:511–524
Kraus P, Leong G, Tan V et al (2010) A more cost effective and rapid high percentage germ-line transmitting chimeric mouse generation procedure via microinjection of 2-cell, 4-cell, and 8-cell embryos with ES and iPS cells. Genesis 48:394–399
Acknowledgements
We are grateful to Siew Lan Lim, Sook Peng Yap, Hsiao Yun Chan, Song Jie, Geraldine Leong, Valerie Tan, Jie Wei Goh, and Manickam Arun Kumar for their helping hands as well as sharing insight and expertise along the way.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media New York
About this protocol
Cite this protocol
Kraus, P., Sivakamasundari, V., Xing, X., Lufkin, T. (2014). Generating Mouse Lines for Lineage Tracing and Knockout Studies. In: Singh, S., Coppola, V. (eds) Mouse Genetics. Methods in Molecular Biology, vol 1194. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1215-5_3
Download citation
DOI: https://doi.org/10.1007/978-1-4939-1215-5_3
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-1214-8
Online ISBN: 978-1-4939-1215-5
eBook Packages: Springer Protocols