Abstract
Current transgenic technologies for gene transfer into the germline of mammals cause a random integration of exogenous naked DNA into the host genome that can generate undesirable position effects as well as insertional mutations. The vectors used to generate transgenic animals are limited by the amount of foreign DNA they can carry. Mammalian artificial chromosomes have large DNA-carrying capacity and ability to replicate in parallel with, but without integration into, the host genome. Hence they are attractive vectors for transgenesis, cellular protein production, and gene therapy applications as well. ES cells mediated chromosome transfer by conventional blastocyst injection has a limitation in unpredictable germline transmission. The demonstrated protocol of laser-assisted microinjection of artificial chromosome containing ES cells into eight-cell mouse embryos protocol described here can solve the problem for faster production of germline transchromosomic mice.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Monteith DP, Leung JD, Borowski AH, Co DO, Praznovszky T, Jirik FR, et al. (2004) Pronuclear microinjection of purified artificial chromosomes for generation of transgenic mice: pick-and-inject technique. Methods Mol. Biol. 240: 227–242.
Hernandez, D., Mee, P.J., Martin, J.E., Tybulewicz, V.L.J., and Fisher, E.M.C. (1999) Transchromosomal mouse embryonic stem cell lines and chimeric mice that contain freely segregating segments of human chromosome 21. Hum. Mol. Genet. 8: 923–933.
Shen, M.H., Mee, P.J., Nichols, J., Yang, J., Brook, F., Gardner, R.L., Smith, A.G., and Brown, W.R. (2000) A structurally defined mini-chromosome vector for the mouse germ line. Curr. Biol. 10: 31–34.
Tomizuka, K., Yoshida, H., Uejima, H., Kugoh, H., Sato, K., Ohguma, A., Hayasaka, M., Hanaoka, K., Oshimura, M., and Ishida, I. (1997) Functional expression and germline transmission of a human chromosome fragment in chimaeric mice. Nat. Genet. 16: 133–143.
Tomizuka, K., Shinohara, T., Yoshida, H., Uejima, H., Ohguma, A., Tanaka, S., Sato, K., Oshimura, M., and Ishida, I. (2000) Double trans-chromosomic mice: Maintenance of two individual human chromosome fragments containing Ig heavy and kappa loci and expression of fully human antibodies. Proc. Natl. Acad. Sci. 97: 722–727.
Co, D.O., Borowski, A.H., Leung, J.D., van der Kaa, K.J., Hengst, S., Platenburg, G.J., Pieper, F.R., Perez, C.F., Jirik, F.R., and Drayer, J.I. (2000) Generation of transgenic mice and germline transmission of a mammalian artificial chromosome introduced into embryos by pronuclear microinjection. Chromosome. Res. 8: 183–191.
Voet T, Vermeesch J, Carens A, Dürr J, Labaere C, Duhamel H, David G, Marynen P. (2001) Efficient male and female germline transmission of a human chromosomal vector in mice. Genome Res. 11(1): 124–36.
Burgoyne, P.S. and Mahadevaiah, S.K. (1993) Unpaired sex chromosomes and gametogenetic failure. In Chromosomes Today, Volume 11 (eds. A.T. Summers and A.C. Chandley) pp. 243–263. Chapman and Hall, New York.
Hunt, P., LeMaire, R., Embury, P., Sheean, L., and Mroz, K. (1995) Analysis of chromosome behavior in intact mammalian oocytes: Monitoring the segregation of a univalent chromosome during female meiosis. Hum. Mol. Genet. 4: 2007–2012.
RamÃrez, M.A., Fernández-González, R., Pérez-Crespo, M., Pericuesta, E., Gutiérrez-Adán A. (2009) Effect of Stem Cell Activation, Culture Media of Manipulated Embryos, and Site of Embryo Transfer in the Production of F0 Embryonic Stem Cell Mice. Biology of Reproduction 80: 1216–1222
Poueymirou W.T., Auerbach W., Frendewey D., et al. (2007) F0 generation mice fully derived from gene-targeted embryonic stem cells allowing immediate phenotypic analyses. Nat Biotechnol. 25: 91–9.
Katona RL, Sinkó I, Holló G, Szucs KS, Praznovszky T, Kereso J, et al. (2008) A combined artificial chromosome-stem cell therapy method in a model experiment aimed at the treatment of Krabbe’s disease in the Twitcher mouse. Cell. Mol. Life Sci. 65(23):3830–3838.
Nagy, A., Gertsenstein, M., Vintersten, K. & Behringer, R.. (2003) Manipulating the Mouse Embryo: a Laboratory Manual. 764 p. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Sinko, I., Katona, R.L. (2011). Transfer of Stem Cells Carrying Engineered Chromosomes with XY Clone Laser System. In: Hadlaczky, G. (eds) Mammalian Chromosome Engineering. Methods in Molecular Biology, vol 738. Humana Press. https://doi.org/10.1007/978-1-61779-099-7_13
Download citation
DOI: https://doi.org/10.1007/978-1-61779-099-7_13
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-098-0
Online ISBN: 978-1-61779-099-7
eBook Packages: Springer Protocols